I

PREVALENCE OF MASTITIS AND ASSOCIATED RISK FACTORS IN LACTATING ONE-

HUMPED CAMELS IN WEST POKOT COUNTY, KENYA

A thesis submitted in partial fulfillment of requirements for Masters of Science degree in Veterinary

Epidemiology and Economic (MVEE), University of Nairobi.

Charles Kisa Toroitich, BVM, University Of Nairobi,

Department of Public Health, Pharmacology and Toxicology,

Faculty of Veterinary Medicine,

University of Nairobi

May 2013

II

DECLARATION

This thesis is my original work and has not been presented for a degree in any university.

…………………………………………….. Date …………………………………..

Dr.Charles Kisa Toroitich BVM University of Nairobi

This thesis has been submitted for examination with our approval as the University supervisors.

……………………………………………… Date ………………………………….

Prof. Philip M. Kitala, BVM, MSc, PhD.

Department of Public Health, Pharmacology and Toxicology

………………………………………………..Date …………………………………….

Prof. George K. Gitau, BVM, MSc, PhD.

Department of Clinical Studies

…………………………………………………Date …………………………………….

Dr. George C. Gitao, BVM, MSc, PhD

Department of Veterinary Pathology, Microbiology and Parasitology

III

DEDICATION

This thesis is dedicated to my dear wife:

Mercy Jepkemei Toroitich

And our children: Kigen Tembu Toroitich

Jesaina Tembu Toroitich

Jerono Tembu Toroitich

IV

ACKNOWLEDGEMENTS

I wish to thank most sincerely all my three supervisors, Profs. Philip M. Kitala, George K. Gitau and

Dr. George C. Gitao for their guidance, advice, encouragement, moral support, patience and

constructive criticism throughout this study.

Special thanks goes to the Coordinator ASAL-Based Livestock and Livelihood Support Project

(ALLPRO) Mr. James K. Tendwa for the financial support during training and collection of data in the

field. I say big thank you Mr. Coordinator.

I also express my sincere gratitude to the District Livestock Production Officer (DLPO) West Pokot

Mr. Andrew Kaptalai and his officer Musa Lelterit, his driver Martin Wesonga, West Pokot DVO’s

Officers Monica Mbithi and Kashmir Magal for their willingness to support in the field. These officers

volunteered their time even weekends to be in the field under the harsh environment of West Pokot

County. Sincere thanks go to Henry Tingoria and Simon Akasilee who were the KCA North Rift

Regional officials for mobilizing the camel keepers and translating from Pokot language to English.

I am indebted to my colleague at Kenya Camel Association (KCA) Head Quarters Mr. Khalif Abey for

the many supports inside and outside the office he assisted me during the study.

This study would not have succeeded without the many roles the camel keepers and herders played in

allowing access to their camels. They provided the answers to the questionnaires and any other

relevant information required.

Finally I thank my family members and great friends of my family for their moral support, continuous

prayers and encouragements during the course of the study.

V

TABLE OF CONTENTS

DECLARATION II

DEDICATION III

ACKNOWLEDGEMENTS IV

TABLE OF CONTENTS V

LIST OF TABLES IX

LIST OF FIGURES XI

LIST OF APPENDICES XII

ABBREVIATIONS AND ACRONYMS XIII

ABSTRACT XIV

CHAPTER ONE 1

1.1 INTRODUCTION 1

1.1.1 Classification and geographical distribution of camels in the world 1

1.1.2 Camel population 2

1.1.3 Local breeds of camels 3

1.1.4 Socio-Economic importance of the camel in Kenya 4

1.2 Problem Statement 7

CHAPTER TWO 9

2.1 LITERATURE REVIEW 9

2.1.1 Introduction 9

2.1.2 Epidemiology of Mastitis 11

2.1.2.1 Occurrence and distribution of mastitis 11

2.1.2.2 Disease determinants for mastitis infections 14

2.1.2.2.1 Disease causative agents 14

2.1.2.2.2 Host factors associated with mastitis 15

VI

2.1.2.2.3 Environmental factors associated with mastitis 16

2.1.2.3 Diagnosis of mastitis 16

2.1.2.4 Treatment of mastitis in camels 18

2.1.2.5 Control of Camel Mastitis 20

2.1.2.6 Economic importance of mastitis in camels 21

CHAPTER THREE 22

3.1 MATERIALS AND METHODS 22

3.1.1 Study area 22

3.1.2 Study Design 26

3.1.3 Selection of the study sites, households and camels 27

3.1.4 Data collection 27

3.1.5 Sample size determination for the number of milk samples 28

3.1.6 Collection of milk samples 29

3.1.7 Examination for sub-clinical mastitis at household level 29

3.1.8 Bacteriological Examination and Isolations 30

3.1.9 Statistical Data Analysis 31

3.1.10 Definition of outcome variables 32

CHAPTER FOUR 33

4.1 RESULTS 33

4.1.1 Characterization of camel management practices 33

4.1.2 Household demographics 37

4.1.2.1 Age and sex distribution 37

4.1.3 Laboratory Results on bacterial culture and identification 39

4.1.3.1 Prevalence of subclinical mastitis at animal (camel) level in the study area 39

4.1.5.1 Bacterial Isolation per division 45

VII

4.2 Results per Division 46

4.2.1 Kongelai Division 46

4.2.1.1 Herd Structure by age of camel in Kongelai division 46

4.2.1.3 Prevalence of subclinical mastitis at quarter level in Kongelai division 48

4.2.2 Kacheliba Division 49

4.2.2.1 Herd structure by age of camel in Kacheliba division 49

4.2.2.2 Prevalence of subclinical mastitis at animal (camel) level in Kacheliba division 50

4.2.2.3 Prevalence of subclinical mastitis at quarter level in Kacheliba division 51

4.2.3 Konyao Division 52

4.2.3.1 Herd structure by age of camels in Konyao division 52

4.2.3.2 Prevalence of subclinical mastitis at animal (camel) level in Konyao division 53

4.2.3.3 Prevalence of subclinical mastitis at quarter level in Konyao division 53

4.2.4 Kiwawa Division 54

4.2.4.1 Herd structure by age of camel in Kiwawa division 54

4.2.4.2 Prevalence of subclinical mastitis at animal (camel) level in Kiwawa division 55

4.2.4.3 Prevalence of subclinical mastitis at quarter level in Kiwawa division 55

4.2.5 Kasei Division 57

4.2.5.1 Herd structure in Kasei division 57

4.2.5.2 Prevalence of subclinical mastitis at animal (camel) level Kasei division 58

4.2.5.3 Prevalence of subclinical mastitis at quarter level Kasei division 59

Alale Division 60

4.2.6.1 Herd structure Alale division 60

4.2.6.2 Prevalence of subclinical mastitis at animal (camel) level Alale division 61

4.2.6.3 Prevalence of subclinical mastitis at quarter level in Alale division 61

CHAPTER FIVE 63

VIII

5.1 DISCUSSION 63

CHAPTER SIX 67

6.1 CONCLUSSIONS AND RECOMMENDATIONS 67

6.1.1 CONCLUSSIONS 67

6.1.2 RECOMMENDATIONS 68

8.1 APPENDIX I: A CROSS SECTIONAL STUDY QUESTIONNAIRE 86

8.2 APPENIX II: Laboratory Results on bacterial culture and identification 92

IX

LIST OF TABLES

Table 3.1: Estimated livestock population in West Pokot County 26

Table 4.1: Camel management practices 34

Table 4. 2: Traditional ways of managing camel mastitis in West Pokot County 36

Table 4. 3: Camel diseases and conditions reported by Pokot herders 36

Table 4. 4: Age and sex distribution of the household's heads interviewed 37

Table 4.5: Summary of herd structure per division 39

Table 4.6: Prevalence of subclinical mastitis at animal (camel) level in the study area 40

Table 4.7: Prevalence of subclinical mastitis at quarter level 40

Table 4.8: Prevalence of subclinical mastitis at two quarters (left & right) level 42

Table 4.9: Prevalence of subclinical mastitis at quarter level showing OR & Chi-2 42

Table 4.10: Association between the occurrence of mastitis and stage of lactation 43

Table 4.11: Association between the occurrence of mastitis and parity 43

Table 4.12: Bacterial isolation rates and their prevalences 44

Table 4.13: Bacterial Isolation per division 45

Table 4.14: Herd structure by age of camel in Kongelai division 46

Table 4.15: Prevalence of subclinical mastitis at quarter level in Kongelai division 48

Table 4.16: Herd structure by age of camel Kacheliba division 49

Table 4. 17: Prevalence of subclinical mastitis at quarter level Kacheliba division 51

Table 4.18: Herd structure by age of camels in Konyao division 52

Table 4. 19: Prevalence of subclinical mastitis at quarter level in Konyao division 53

Table 4.20: Herd structure Kiwawa division 54

Table 4. 21: Prevalence of subclinical mastitis at quarter level in Kiwawa division 56

Table 4.22: Herd structure Kasei division 57

X

Table 4.23: Prevalence of subclinical mastitis at quarter level in Kasei division 59

Table 4.24: Herd structure Alale division 60

Table 4. 25: Prevalence of subclinical mastitis at quarter level in Alale division 62

XI

LIST OF FIGURES

Figure 3.1: Map of Kenya showing West Pokot County the study area 22

Figure 3.2: Map of West Pokot County showing the six study divisions 24

Figure 4.1: Age and sex distribution of the households' heads interviewed 38

Figure 4.2: Prevalence of subclinical mastitis per quarter 41

Figure 4.3: Herd structure in Kongelai division 47

Figure 4. 4: Herd structure Kacheliba division of West Pokot County 50

Figure 4.5: Herd structure Konyao division of West Pokot County 52

Figure 4.6: Herd structure in Kiwawa division of West Pokot County 55

Figure 4.7: Herd structure in Kasei division of West Pokot County 58

Figure 4.8: Herd structure Alale division of West Pokot County 61

XII

LIST OF APPENDICES

Appendix 1: a cross sectional study questionnaire 80

Appendix II: Laboratory Results on bacterial culture and identification 95

Appendix III: Photographs from the field and in the Laboratory 101

XIII

ABBREVIATIONS AND ACRONYMS

ALLPRO ASAL Based Livestock & Livelihood Support Project

ASAL Arid and Semi Arid Lands

BA Blood Agar

CAMP Christie, Atkins and Munch-Petersen

Chi-2 Chi-square

CMT California Mastitis Test

CPP Country Program Paper

DNA Deoxyribonucleic Acid

FAO Food and Agriculture Organization of the United Nations

HH Households

HoA Horn of Africa

KCA Kenya Camel Association

MA MacConkey Agar

LFQ Left Fore Quarter

LHQ Left Hind Quarter

OR Odds Ratio

RFQ Right Fore Quarters

RHQ Right Hind Quarter

STAT Statistics

Spp Species

XIV

ABSTRACT

In spite of it living in harsh environments of semiarid and arid zones, the dromedary camel is

able to produce milk in valuable quantity. Camel milk is one of the main components of diet of the

nomads in semiarid and arid zones and is an essential food for livelihood of people and it may be the

only milk available in the ASALs where other milking animals cannot be maintained. However, like

other dairy animals, dromedary camels could be affected by udder infections such as mastitis, a

complex disease occurring worldwide among dairy animals, with heavy economic losses largely due

to clinical and subclinical mastitis.

A cross sectional study was conducted to determine the prevalence of mastitis and to identify

the associated risk factors in 95 clinically healthy lactating and traditionally managed one-hump

camels (Camelus dromedarius) in Kongelai, Kacheliba, Konyao, Kasei, Kiwawa and Alale divisions

of West Pokot County, Kenya. Fifty two households were conveniently selected from a list provided

by Kenya Camel Association West Pokot County based on the presence of a lactating camel in the

household. Data on camel management including milking procedures were collected through

interviews using closed ended questionnaires.

A total of 380 quarter milk samples (56 from Kongelai division, 40 from Kacheliba division, 8

from Konyao division, 148 from Kiwawa division, 92 from Kasei division and 36 from Alale division)

were collected aseptically. The samples were transported in cool boxes with ice packs to the

Bacteriology Laboratory at the Department of Veterinary Pathology, Microbiology and Parasitology,

Faculty of Veterinary Medicine, University of Nairobi for bacterial culture. Of the 380 quarter milk

samples cultured, 169 samples tested positive for subclinical mastitis which gave a prevalence of

quarter infection at 44.5% (169/380). At animal (camel) level the prevalence of subclinical mastitis

was 76.8% (73/95 camels).Therefore the results of this study showed that subclinical mastitis is

prevalent in dromedary camels of West Pokot County. The same results showed that, the right hind

XV

quarter (RHQ) was the most frequently infected quarter (prevalence of 12.1% (46/380)) followed by

the right fore quarter (RFQ) (prevalence of 11.3% (43/380)). The two left quarters, left fore quarter

(LFQ) &left hind quarter (LHQ) were least infected. This could point out that the Pokot herders tended

to milk the right quarters more often and left the left quarters to be suckled by the calves and because

of poor & unhygienic milking procedures the right quarters become more infected.

The most predominant isolated bacterium was gram-positive Staphylococcus aureus with

prevalence of 36.0% (49/136) followed by gram-negative Escherichia coli with prevalence of 27.2%

(37/136). Streptococcus agalactiae & Staphylococcus epidermidis were the third predominant isolates

with prevalence of 9.6% (13/136) each. Micrococcus spp & Pseudomonas were least isolated with less

than 1% prevalence each. A diagnosis of ‘no bacterial growth’ was made in 22 cases; which translates

to 16.2% (22/136). Several mastitis control strategies need to be put in place such as milking

procedures, milking order, strict hygiene, post milking teat disinfection, use of antibiotic dry-off

therapy and the culling of persistently infected camels.

Significant (p<0.05) differences in subclinical mastitis prevalence were observed between

camels in different lactating stages and parities. Camels in more than two months lactation stages were

affected at higher rate (OR=2.75, p<0.05) than those in less than two months lactation stages. Also

camels which had given birth to more than two calves (second parity or more) were affected at higher

rate (OR=2.90, p<0.05) compared to camels which have given birth to less than two calves.

The fact that the pathogens isolated from camel milk samples in this study were bacteria that cause

both environmental and contagious mastitis, this study concludes that proper management of lactating

camels and adequate hygienic conditions of the environment are required in order to minimize

occurrence of mastitis in the study areas. It also recommends treatments of camels with mastitis

infections using the conventional drugs and avoid non-conventional treatment.

Keywords: Dromedary camels, subclinical mastitis, quarter samples, West Pokot Kenya.

1

CHAPTER ONE

1.1 INTRODUCTION

1.1.1 Classification and geographical distribution of camels in the world

In zoological taxonomy, Camelids are classified in the suborder Tylopoda (pad-footed animals)

that represents with the suborders Suiformes (pig-like) and Ruminantia (ruminants) the order

Artiodactyla (even-toed ungulates). This makes obvious that Camelids (family Camelidae) as

ruminating animals are classified in proximity to ruminants but developed in parallel and are not

part of the suborder Ruminantia. Some differences like foot anatomy, stomach system and the

absence of horns underline this fact (Schwartz & Dioli, 1992; Fowler, 1998; Wernery, 2003).

The family Camelidae is divided into three genera: The old world camels (genus Camelus) and

the new world camels (genus Lama with the species L. glama, L. guanicoe, L. pacos and genus

Vicugna with the species V. vicugna) (Wilson & Reeder, 2005). Two domesticated species of old

world camels exist: the dromedary or one humped camel (Camelus dromedarius,) that has its

distribution in the hot deserts of Africa and Asia and the Bactrian or two-humped camel

(Camelus bactrianus) that can be found in the cold deserts and dry steppes of Asia. In the desert

Gobi there is still a population of wild two-humped camels classified as Camelus ferus (Rao et

al., 1970; Peters, 1997; Fowler, 1998).

The Bactrian camel was named after the area of Bactriana in Central Asia. The name of the

dromedary was derived from the Greek word “dromeus” which means runner or “droma” -

running (Jassim & Naji, 2002). The one-humped camel was probably domesticated in the region

of today’s Yemen and Oman about 3.000 to 4.000 years ago (Fowler, 1998). The wild Arabian

2

camel became extinct (Lensch, 1999). Curasson (1947) and Epstein (1971) indicate that the

dromedary (Camelus dromedarius) was introduced into North Africa (Egypt) from Southwest

Asia (Arabia and Persia). The former indicates that occasional shipments were also made to

Spain, Italy, Turkey, France, the Canaries, North America and Australia. The latter country still

contains a small feral herd of around 20,000. Once in Africa, Mikesell (1955) suggests that the

camel spread West and Southwards from Egypt, although Bulliet (1975) is of the view that the

camels of the Horn of Africa are more likely to have come across the sea from the Arabian

Peninsula than spread southwards from Egypt and Sudan.

In East Africa, it is thought that the camel was introduced following a more direct route through

the Horn of Africa during the middle of the 1st millennium BC (Epstein 1971). The camels found

their way to Kenya from Somalia after domestication in Southern Arabia between 1 & 4 B.C

(Bulliet, 1975; Wilson, 1984).

1.1.2 Camel population

According to FAO statistics (Global Livestock Production and Health Atlas - GLIPHA, 2006)

the world population of camels is about 20 million, mainly in arid zones, of which 15 million

live in Africa and 5 million in Asia (GLIPHA, 2006). In 2001, the total camel population was 19

million of which 17 million were dromedaries (Camelus dromedarius) and 2 million were

Bactrian camels (Camelus bactrianus) (Farah, 2004). In most countries, the camel population is

increasing after a period of decreasing number due to the introduction of modern transport

facilities (Farah, 2004). Kenya has an estimated dromedary camel population of 2,971,000, with

majority of the camels about 1,700,000 (57%) in North Eastern followed by Rift Valley Province

carrying 968,000 (33%) camels (Kenya National Census, 2009). West Pokot County has a

3

population of 30,600 camels with majority of the camels found in Pokot North Sub County

(Kenya National Census, 2009).

1.1.3 Local breeds of camels

There are three local recognizable dromedary camel breeds in Kenya which are named after the

pastoral communities who own and keep them. These are Somali, Rendille/Gabbra and Turkana

breeds; the former generally being the largest and the later the smallest in size (Bremaud, 1969,

Simpkin, 1983). The fourth breed is the exotic “Pakistani” breed which was introduced into

Kenya in the last three decades by researchers and development agents (Hulsebusch and

Kaufmann, 2002). This breed has better productive and genetic characteristics compared to the

local breeds.

The Somali breed camels are named after the pastoralist group of the same name and referred to

as Benadir camels (probably the same as the Benadir type found in Somalia) by some authors

(Wilson, 1984). They are primarily owned by the Somali people of North-Eastern province of

Kenya, are the most productive local breed, and can be considered as part of the larger

population of camels in Somalia (Karue, 1989). This breed of camels is generally larger than the

other breeds found in the country (Bremaud, 1969; Kegode, 1990). Field (1993) and Simpkin

(1995a) reported that Somali breed camel owners in Kenya differentiated camels into three, or

sometimes four, races: that is Sifdaar, Hoor, Gelab and Aidimo.

These different races may be associated to certain Somali clans or families (Simpkin, 1995a).

According to Simpkin (1996) the Somali breed camel in Kenya produces more milk than the

Turkana breed under the identical conditions. Rendille/Gabbra breed camels are found mainly in

Marsabit County amongst the Rendille and Gabbra tribes (Simpkin, 1995a). The traditions of

4

these people against selling or trading breeding females with other people have limited the

distribution of these camels (Stiles, 1995). The Turkana breed camels are commonly found in

Turkana County as well as Samburu and Pokot counties where they were obtained by trading or

raiding (Stiles, 1995).

1.1.4 Socio-Economic importance of the camel in Kenya

The ASALs occupy 89% of the of the Kenyan landmass of which 70% is arid (Northern Kenya)

and 19% semi-arid lands dispersed all over the country (Government of Kenya Sessional Paper

No 8 for 2012 and Kenya Country Program Paper (CPP) October 2012). The ASALs are home to

about 14 million people, of whom 4 million are pastoralists (Kirbride and Grahn 2008).

Approximately 95% of ASAL households derive their income from the livestock subsector

where 70% of livestock is produced. The camel is considered to be potentially the most

important animal source of food in pastoral areas (Farah et al., 2006).

Schwartz and Dioli, (1992) reported that dromedary camel is a multipurpose animal adapted to

the harsh environments of semi-arid and arid zones, essentially kept for milk and meat

production and transportation. It is also a financial reserve (asset) and security (drought-prone

risk management) for pastoralists and plays an important role in social prestige and wealth.

The position of the camel in providing food for the pastoralists in Northern Kenya may become

even more important in the face of global warming and climate change ( Ndikumana et al,.

2000). Camels (Camelus dromedarius) are multipurpose animals increasingly kept for milk and

meat (Abdurahman, 2005). Nomadic pastoralist communities living in ASAL regions largely

5

depend on milk produced by camels which contribute 80% of the household needs (Schwartz and

Dioli, 1992; Guliye, 2006).

Camel milk is one of the main components of diet of the nomads in semiarid and arid zones and

is an essential food for livelihood of people and it may be the only milk available in places where

other milking animals cannot be maintained (Kazmi, 2002; Abdurahman, 2006). Camels may

produce six times the volume of milk produced by local cattle under the same arid conditions

(Field and Simpkin, 1985). This milk is an important source of protein (Yagoub, 2003) for

nomadic tribes and the ignored rural citizens. Also it is a good source of vitamin C in these areas

where the traditional sources of vitamin C are rare (Schwartz and Diole, 1992; Wilson, 1998).

The amount of vitamin C in camel milk is said to be three times more than in the cow’s milk,

iron content ten times and B vitamins present in reasonable amounts (Barbour et al., 1985;

Elagamy et al., 1992; Arrowal et al., 2005). Unfortunately, many reports revealed that lactating

she-camels easily succumb to mastitis (Abdurhman et al., 1995, Obied et al.,1996, Abdel Gadir

et al., 2006).

Apart from being source of food, camels’ milk is also taken traditionally for the control

and management of diabetes type-1 and a recent study in India has given scientific support to this

belief (Agrawal et al., 2002). Clinical trials in human diabetes mellitus type 1 have shown that

camel milk reduces the need for insulin medication by an average of 30% (Agrawal et al., 2005).

This is attributed to the fact that camels browse on various plant species and the active agents

with therapeutic properties from these plants are secreted into the milk of camels (Muli et al.,

2008). There is also an account in the memories of Emperor Jahangir (1579-1627 AD) about the

usefulness and acceptability of camel milk (Rogers, 1989). It was found that one of the camel

milk proteins has many characteristics similar to insulin (Beg et al, 1989). Furthermore, it does

6

not form a coagulum in an acidic environment (Wangoh, 1993). Oral insulin therapy has been

known for many years but the important drawback is its coagulum formation in acidic

environment such as the stomach, thereby neutralizing its potency. This lack of coagulum

formation allows the camel milk to pass rapidly through the stomach together with the specific

insulin like-protein and remains available for absorption in the intestine. Radioimmunoassay

tests of camel milk has revealed high concentration of insulin at about 52 micro units/ml. The

concentration of insulin in human milk is also significantly higher (60.23 ± 41.05 micro units/ml)

whereas it is low in cow milk (16.32 ± 5.98 micro units/ml) (Shehadeh et al, 2001). There is

strong evidence that oral insulin products would provide insulin in a more physiological manner,

resulting in a decrease in peripheral insulin concentrations thus “insulinsing” life (Gwinup et al,

1991 and Hoffman and Siv, 1997).

The camel is considered the most important dairy animal in Kenya’s Arid and Semi-Arid

Lands (ASALs) and according to Muli et al. (2008), camel milk production in Kenya in 2007

was estimated to have stood at over 340 million litres. Only about 12% of the milk was

marketed, the bulk of which was sold in raw form to rural consumers (10%) and only 2%

reached urban consumers. From the remaining milk (88%) that did not reach the market, 38%

was directly used by camel keeping households and their herders as part of their food

requirements and the remaining 50% (or 170 million litres) went into waste representing a great

opportunity for commercialization and enhanced incomes for communities in pastoral areas.

Mastitis has both an extreme zoonotic and economic importance and it is the cause of

multiple hazardous effects on human health and animal production (Makovec and Ruegg, 2003;

Hegazy et al., 2004; Al-Majali et al., 2008). Little work has been done on mastitis in camels

comparing to studies on sheep and cows. Three decades ago, there was no mention of mastitis

7

problem at herd level; today it is reported from almost all camel rearing countries (Al-Ani and

Al-Shareefi, 1998; Guliye et al., 2002; Khedid et al., 2003; Mohammed et al., 2005). Milk is a

nutritious food for human beings, but it also serves as a good medium for the growth of many

microorganisms, especially bacterial pathogens. Lactococcus, Lactobacillus, Streptococcus,

Staphylococcus and Micrococcus spp. are among the common bacterial flora of fresh milk (Chye

et al., 2004).

Many different bacteria have been isolated from mastitic mammary glands in camels either in the

form of pure or mixed infection (Abdel Gadir et al., 2006; Abdurahman, 2006; Hegazy et al.,

2004; Bekele and Molla, 2001;Woubit et al., 2001; Younan et al., 2001; Abdurahman, 1996 and

Barbour et al., 1985).

1.2 Problem Statement

Camels are adapted to the ASALs, but their full milking potential is affected by udder infections

especially sub-clinical mastitis and yet little work has been done on mastitis in camels compared

to studies on cows, goats and sheep.

8

1.3 Objectives

The overall objective of this study was to estimate the prevalence of mastitis and the associated

risk factors in traditionally managed one-humped camels in West Pokot County of Kenya. This

main objective was achieved through the following two specific objectives;

a) To estimate the prevalence of clinical and sub-clinical mastitis in lactating camels in

West Pokot County.

b) To determine the potential risk factors associated with the prevalence of mastitis in

lactating camels in the study County.

9

CHAPTER TWO

2.1 LITERATURE REVIEW

2.1.1 Introduction

In spite of it living in harsh environments of semiarid and arid zones, the dromedary

camel is able to produce milk in valuable quantity (Schwartz and Dioli, 1992; Faye, 2005).

However, like other dairy animals, dromedary camels could be affected by udder infections such

as mastitis, a complex disease occurring worldwide among dairy animals, with heavy economic

losses largely due to clinical and subclinical mastitis. The latter requires indirect means of

diagnosis (Matofari et al., 2003). Camel mastitis is both medically and economically important

due to its multiple hazardous effects on human health and animal production (Younan, 2004;

Akweya et al., 2010; Njage et al., 2010). In addition to these health concerns, mastitis reduces

production (Musinga et al., 2008) and quality (Matofari et al., 2003; Mengistu et al., 2010) of

milk of traditionally managed camels. In the arid and the semi-arid areas of Kenya (ASALs),

milk is consumed fresh or sour posing a health hazard to consumers (Younan, 2004). It has

recently come to light (Younan et al., 2001) that infections of the udder of lactating camels are

quite widespread. Generally, bacteria in milk can occur through colonization of the teat canal or

an infected udder (clinical or subclinical mastitis), or as contaminants (Younan, 2004).

Mastitis can be defined as the inflammation of the mammary gland regardless of the

cause and is characterized by physical, chemical and, usually, bacteriological changes in the

milk. It is also characterized by pathological changes in the glandular tissue. The most important

changes in the milk include discoloration, the presence of milk clots and the presence of a large

10

number of leucocytes (Radostits et al, 2000). Although there is supposed to be swelling, heat,

pain and indurations in the mammary gland, a large proportion of mastitis cases are not readily

detectable by manual palpation, neither by visual examination of the milk using a strip cup; such

cases are referred to as “subclinical mastitis”. Because of the very large number of sub-clinical

mastitis cases, the diagnosis of mastitis has become dependent largely on indirect tests which

depend in turn on the leucocytes content of the milk (Radostits et al., 2000).

Evidence indicates that subclinical mastitis causes suffering of the animal, reduces milk

yield, alters milk properties, impairs preservation and processing and is of public health concern

for consumers of camel milk (Fthenakis and Jones, 1990; Tibary and Anouassi, 2000). Very little

is known about aetiology and occurrence of mastitis in Camelidae (Abdel Gadir et al., 2006;

Kalla et al., 2008). However, cases of mastitis in camel have recently been reported in Saudi

Arabia (Barbour et al., 1985); Egypt (Mostafa et al., 1987); Somalia (Abdurahman et al., 1991);

Ethiopia (Bekele and Molla, 2001); Israel (Guliye et al., 2002) and Kenya (Matofari et al., 2003).

During the past decade there have been several reports on subclinical mastitis in dromedary

camels (Obeid, 1983; Arush et al., 1984; Quandil and Oudar, 1984; Barbour et al., 1985;

Mostafa et al., 1987) and a few in Bactrian camels (Kospakov, 1976a,b); however, little work

has been done on subclinical mastitis and the udder’s response to bacterial invasion. Barbour et

al. (1985) and Saber et al. (2010) applied CMT to composite milk samples from the dromedary

camels and concluded that the test was useful for screening subclinically infected udders. Obeid

(1983) found a good correlation between the milk leukocyte count and the ‘rapid mastitis test’.

The prevalence of mastitis causing organisms in camel milk is a concern of Public Health. Early

problem recognition and improved hygiene can reduce the milk loss due to mastitis resulting in

high economic gain (Abdurahman, 2006). Better herd management of mastitis can also increase

11

milk production and thus income for pastoralists (Abdurahman and Younan, 2004). The greatest

economic loss of mastitis in both subclinical and clinical mastitis is reduced milk production

(Wood and Booth, 1983).

Traditionally, the Pokots are nomadic pastoralists whose lifestyle rotates mainly on

livestock keeping but there is evidence to suggest that they have had a longer involvement with

camels in the past (McGovern, 1995). Their recent acquisition of camel was through their role as

mercenaries during punitive raids against the Turkana in 1917, or trading with Somali camel

traders in the 1950s (Bollig, 1992).

Access of the camel calf to the dam is the most commonly used stimulus for initiating milk let

down (Dorman, 1984). The calves are often only permitted to initiate milk letdown by suckling

for 1-2 minutes (Mares, 1954; Hashi, 1984; Simpkin, 1985) and are then either totally restricted

from suckling whilst all the four quarters are milked, or permitted to suckle one or two teats

whilst the remainder are milked.

2.1.2 Epidemiology of Mastitis

2.1.2.1 Occurrence and distribution of mastitis

The prevalence and causes of mastitis differ markedly due to geographical area and

individual herd management (Guidry, 1985). Even in well-managed herds, as judged by somatic

cell count level and a low level of milk production, there may still be occurrence of high

incidence of clinical mastitis (Erskine et al., 1989; Hogan et al., 1990 and Schukken et al.,

1991).

12

Clinical mastitis is mostly caused by bacteria with the most important causative agents being

Staphylococcus aureus, Streptococcus spp (Streptococcus uberis and dysagalactia), Escherichia

coli and Klebsiella spp.

Clinical mastitis is only an indicator of the herd infection though subclinical mastitis is by far the

more costly disease in the majority of herds, and is often defined as the presence of a

microorganism in combination with an elevated somatic cell count (SCC) in the milk. In

subclinical mastitis, there are no obvious clinical signs such as milk clot and flakes, udder

swelling or tenderness, or systemic signs such as fever, depression. Instead there is an increase in

somatic cell counts of the milk (Radostis et al., 1999).

While clinical mastitis is rather easy to detect, animals suffering from subclinical mastitis

are often very difficult to find since there is lack of reliable diagnostic methods, especially at

farm level (Leitner et al., 2004). However, two indirect tests viz. somatic cell count and bacterial

load count are accepted reliably for detecting the early infection (Schalm et al., 1971). In

addition, other indirect tests like White Side Test and California Mastitis Test (CMT) have been

developed for rapid screening of udder infection (Schalm et al., 1971; Guha et al., 1989). If the

cases are not detected on time, subclinical mastitis may progress and develop into clinical

mastitis (Adwan et al., 2005). Mastitis is a complex disease problem and presents as a classical

example of the interaction of microorganisms, host factors and the environment.

Infection patterns

There are two distinct patterns in the epidemiology of mastitis that can be recognized;

The contagious disease pattern where transfer of microorganisms from animal to animal is

essential to propagate the disease.

13

The opportunistic microorganisms’ pattern where host factors and environmental factors put an

animal at risk. A wide range of microorganisms can then enter the mammary gland and cause the

disease.

Contagious mastitis can be transmitted from an infected or carrier animal to a susceptible host.

The organisms mainly associated with the spread of contagious mastitis in the dairy population

are Streptococcus agalactiae and Staphylococcus aureus (Natzke, 1981). Other epidemic

contagious disease outbreaks have been reported, and involve Nocardia spp, Mycoplasma spp.

and in some situations environmental Streptococci.

Contagious diseases only remain endemic when the mean number of susceptible individuals

infected by the respective organism is appreciably larger than one (Becker, 1989).

Reduction of the number of new mastitis infections is the major goal of any mastitis prevention

program. New mastitis infections may be reduced by optimizing milking procedures and post

milking teat disinfection. These practices can reduce the number of shedders in the herd, separate

the shedders from the uninfected camels, and optimize the immune function of the animal, which

are key components of decreasing new infections. Eliminating existing infections reduces the

exposure of susceptible quarters and may be obtained by treatment during lactation or at dry off,

or by culling of the infected animals. Again, separation of the infected animals from the

susceptible group may also be an effective method to limit the exposure of susceptible animals

and reduce the risk of new infections.

14

2.1.2.2 Disease determinants for mastitis infections

2.1.2.2.1 Disease causative agents

Historically, mastitis pathogens have been classified as either “contagious” or

“environmental” (Blowery & Edmondson, 1995). The contagious pathogens are considered as

organisms adapted to survive within the host, in particular within the mammary gland, and are

typically spread from animal to animal at or around the time of milking (Radostits et al., 1994,

Blowery and Edmondson, 1995). In contrast, the environmental pathogens are best described as

opportunistic invaders of the mammary gland, not especially adapted to survival within the host;

typically they enter, multiply, elicit a host immune response and are eliminated. The major

contagious pathogens are Staphylococcus aureus, Streptococcus dysgalactiae and Streptococcus

agalactiae and the major environmental pathogens are the Enterobacteriacae and Streptococcus

uberis.

Incidences of pathogenic organisms in camel milk have been reported in Kenya (Younan

et al., 2000). The most common isolates from camel mastitis are Streptococcus agalactiae and

Staphylococcus aureus, however other isolates such as Streptococcus uberis, Streptococcus

dysgalactiae, Streptococcus pyogenes, Diplococcus pneumonia, Escherichia coli, Bacillus

cereus, Corynebacterium bovis, Pseudomonas aeruginosa, Pasteurella spp., Pasteurella

haemolytica (chronic suppurative mastitis), Klebsiella spp., Corynebacterium

pseudotuberculosis, Corynebacterium equi and Corynebacterium Pyogenes, Candida albicans

have also been reported (Barbour et al., 1985; Almaw and Molla, 2000 and Bekele and Molla,

2001).

15

Smith et al., (1985) reported that the most important microorganisms involved in mastitis are the

coliforms (E. coli and Klebsiella spp.) and the environmental Streptococcus spp. The severity of

clinical mastitis depends on other microorganism related factors such as serum resistance and

antigen determinants. However, any control/treatment approach that is only based on

microorganism elimination is likely to fail because most host and environmental risk factors will

still remain. It is likely that another microorganism will fill the niche that is created by expelling

one specific organism. Many different bacteria have been isolated from mastitic mammary

glands in camels either in the form of pure or mixed infection (Barbour et al., 1985;

Abdurahman, 1996; Bekele and Molla, 2001; Younan et al., 2001; Woubit et al., 2001; Hegazy

et al., 2004; Abdurahman, 2006 and Abdel Gadir et al., 2006). The prevalence and causes of

mastitis differ markedly due to geographical area and individual herd management (Guidry,

1985).

2.1.2.2.2 Host factors associated with mastitis

Several host factors are important in determining the probability of an infection in camels.

Most infections do not result in the development of clinical signs, and host characteristics’ for

example: peripheral blood leukocyte activity, blood leukocyte count, and presence of antibodies

partially predict the outcome of infection (Lohuis, 1989). Other factors such as age of the animal,

nutritional status, stress and milk production level also affect the outcome of infection. Animals

in early lactation appear to be more susceptible to clinical mastitis and have a relatively high

probability of becoming severely sick.

16

2.1.2.2.3 Environmental factors associated with mastitis

Environmental factors play significant role in the prevalence of sub-clinical (Sandholm,

1995) and clinical (Honkanen-Buzalski and Pyörälä, 1995) mastitis in dairy animals. Several

factors in the environment affect the exposure of a camel to microorganisms. Sources of

environmental contamination include manure, milkers hands, feeds, dirt, mud and water. A good

example of this is E. coli, which is present in the environment of the camel.

Observational studies have shown that most infections with coliform and environmental

Streptococcus take place in the last two weeks before calving, and often only show signs of

clinical mastitis after calving. Reducing exposure of the mammary gland by improving hygiene

or providing a physical barrier at the teat end have shown to reduce the incidence of mastitis.

2.1.2.3 Diagnosis of mastitis

a) California mastitis test (CMT)

Schukken et al. (1988) reported that CMT remains the only reliable screening test for

detection of subclinical mastitis in dairy herds. Kapaga et al. (1995) concluded that CMT test is a

good tool for epidemiological survey of sub-clinical mastitis in dairy herds. Abdurahman (1998)

stated that 100 % of the CMT positive samples tested were positive with pathogenic bacteria.

Apparently there is a significant positive correlation between positive CMT results and the

presence of clinical mastitis in dromedaries (Barbour et al., 1985; Kinne & Wernery, 2002). This

leads to the presumption that the camel, like the cow, has phagocytic cells as one of the essential

defence mechanisms of the mammary gland against pathogenic microorganisms (Schalm et al.,

1971; Barbour et al., 1985; Abdurahman et al., 1992; Saad & Thabet, 1993).

17

b) Electrical activity in camel milk.

The electrical conductivity was not considered adequate as method for mastitis diagnosis

in camels by Younan et al. (2001) and Bhatt et al. (2004) as no significant change can be proved

in case of mastitis. Electrical conductivity is defined as the resistance of a material to electric

current. It is widely used as a simple and effective tool for mastitis diagnosis in cows. In case of

mastitis, the cell membranes of the udder parenchyma are damaged. This increases the

permeability of the barrier between blood and milk. The content of chloride (Cl-) and sodium

(Na+) increases and the content of lactose and potassium (K+) decreases leading to a higher

electrical conductivity of the milk. The average conductivity of cow milk ranges between 4 and

5.8 mS/cm and depends on lactation stage, age, milking interval and race of the individual

animal (Nielen et al., 1992; Walzel, 1997; Billon et al., 2001).

c) N-acetyl-ß-D-Glucosaminidase (NAGase)

N-acetyl-ß-D-Glucosaminidase (NAGase) is a lysosomal enzyme released from damaged

epithelial and other somatic cells in the mammary gland estimated as a good indicator of mastitis

in bovine and ovine milk. In camel milk NAGase activity is significantly higher than in cow milk

which might be due to the high count of cell fragments. It does not clearly correlate with

bacterial findings in contrary to somatic cells count (SCC) (Abdurahman, 1995; Guliye, 1996;

Chaffer et al., 2000). Therefore NAGase activity has not been investigated as a diagnostic mean

for infections or inflammations of camel udders.

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d) Further diagnostic means for camel mastitis

Billon et al. (2001) proposed as additional diagnostic mean of subclinical mastitis in

cows the close observation of the daily milk yield. In camels, the implementation of this idea

could be difficult as dromedaries react very sensitively to their environment with yield

variations. But generally, it is important to verify the udder health and general health status of the

camels. As camels are very sensitive to udder pain, development of clinical mastitis can be easily

detected.

2.1.2.4 Treatment of mastitis in camels

Various procedures have been proposed and/or tried for mastitis treatment in camels.

Although some authors have suggested daily intramammary infusions with an antibiotic

preparation, as applied in cows, there is reservation to this practice because of the particular

anatomy of the camelidae udder and because of the difficulty in administering such treatment

(Tibary and Anouassi, 2000). The therapeutic approach in treating acute mastitis is via systemic

antibiotics (e.g. trimethoprim – sulfamethoxazole or penicillin/ Aminoglycoside) and anti-

inflammatory drugs (flunixin meglumine), with regular stripping of the mammary glands.

Hydrotherapy is also beneficial in reducing local edema.

Due to the smaller diameter of camel teats, intramammary tubes designed for

administration in cattle are often unsuitable for routine use in camels. Currently, antibiotic

intramammary tubes have been of limited success and it may be necessary to design an

applicator with a finer nozzle (Younan, 2002). Camel teats have two or three teat orifices. There

19

is still lack of consensus as to whether the different teat openings also represent separate gland

complexes (Khanna , 1986; Wernery , 2003) .

The teat of the camel udder may sometimes contain three separate teat canals that open

independently into the teat sphincter. The separate canals drain separate gland complexes

(Nosier, 1974); Smuts and Bezuidenhout, 1987). The latter implies that for intramammary

treatment of mastitis, not only must each quarter but also each gland complex be treated

separately, that is, one intramammary tube per gland complex. Great caution is therefore

necessary when applying intramammary treatment to camels given that the teat canal openings in

camel are smaller than those of the cow and thus require smaller canula. Unhygienic and

traumatic application of intramammary treatment is very likely to cause more harm than good.

In the absence of control measures, Streptococcus agalactiae is the most common

mastitis pathogen in dairy cattle (Aguilera, 1984) with average morbidity rates of 25% (Radostits

et al, 1997). Streptococcus agalactiae eradication programs have been successful in dairy cattle

herds and are economically justifiable (Edmondson, 1989); Hejlicek, 1994; Radostits et al,

1997). Intramammary infections (IMI) with Streptococcus agalactiae (Lancefield type B) in

camels are common and have been diagnosed in the United Arab Emirates (Quandil and Qudar,

1984); Egypt (Karamy, 1990); Sudan (Abdurahman et al, 1995; Obied et al., 1996) and Somalia

(Younan et al, 2002). In Northern Kenya, Streptococcus agalactiae IMI prevalence of up to 50%

in market oriented camel dairy herd (Younan et al, 2001) have become a concern to camel

owners. One case of successful parenteral treatment of mastitis in a camel is reported in the

literature (Barbour et al., 1985). However, published treatment recommendations for mastitis in

camels have not been validated (Youssef, 1992; Faye, 1997).

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Pastoralists also use various traditional (ethno-veterinary) practices to treat sick camels

(Bornstein, 1993; Hussein, 1993). However, the success and efficacy of these methods have not

been verified.

2.1.2.5 Control of Camel Mastitis

The specific steps of all udder health management programs must be devised to fulfill

three basic principles and these are: - elimination of existing infections, prevention of new

infections and monitoring of udder health status (Radostits et al., 1994). To establish an efficient

mastitis control program in a dairy herd, baseline information on the nature of mastitis and

economic impact of the problem needs to be known (Honkanen-Buzalski and Pyörälä, 1995).

The principal steps in mastitis control program are to undertake a preliminary mastitis screening

survey and to evaluate the udder health status in the herd (Honkanen-Buzalski and Pyörälä,

1995).

Mastitis can be prevented or reduced by improving animal health and udder hygiene but

currently, there appears to be non-existence of modern mastitis control measures practiced by

camel keepers. Attention must be paid to udder health and hygiene, not only during lactation,

but even when the animal is dry. Animals suffering from any contagious disease, including

mastitis, should be separated from the healthy animals and milk from diseased camels should be

kept separate and disposed off safely. It is cheaper and easier to prevent mastitis by improving

hygienic measures and culling chronically-infected camels to eliminate important pathogen

reservoirs, than to treat by medication. The cost of treatment includes veterinary fees, medicine,

21

and the risk of quackery and costs of milk production. Any unprofessional management or

treatment of camel mastitis can contribute to the buildup of antibiotic resistance.

2.1.2.6 Economic importance of mastitis in camels

Controlling mastitis is important for the dairy industry because the condition has

significant ramifications. These include financial losses to dairy farmers, adverse effects on dairy

animal welfare and potential influences on public health. The broad use of antibiotics in the

treatment and control of mastitis has possible implications for human health through an increased

risk of antibiotic resistant strains of bacteria emerging that may enter the food chain (White and

McDermott, 2001).

Financial loss from clinical mastitis arises from the costs of treatment, culling, death,

decreased milk production and decreased milk revenue. A single case of clinical mastitis is

associated with average losses of around £175 (Kossaibati, 2000), and clinical mastitis on an

average dairy unit accounts for approximately 38% of the total direct costs of the common

production diseases (Kossaibati & Esslemont, 1997). Clinical mastitis in bovine alone has been

estimated to cost the UK dairy industry in excess of £168 million per annum (Bradley, 2002).

It is more difficult to quantify the losses associated with sub-clinical mastitis, because these are

more variable, but losses arise from treatment costs, reduced milk yield, and decreased

constituent quality, loss of livelihood and an increase in the risk of culling. However there are no

estimated economic estimates available as per now.

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CHAPTER THREE:

3.1 MATERIALS AND METHODS

3.1.1 Study area

The study was carried out in West Pokot County in the former Rift Valley Province of Kenya as

shown in Figure 3.1.

Source: Ministry for development of Northern Kenya and other arid lands Figure 3.1: Map of Kenya showing West Pokot County the study area

West Pokot County is one of the 14 Counties that made the former Rift Valley Province

of Kenya (Constitution of Kenya, 2010). It borders Uganda to the West, Trans Nzoia and

Elgeyo-Marakwet Counties to the South, Turkana County to the North and East and Baringo

23

County to the South East. Geographically, it lies between Latitudes 1010’ and 30 40’ N and

Longitudes 340 50’and 350 50’E (Macmillan Education Ltd., 1999). The County has a total area

estimated at 9,100 square kilometers and stretches a distance of 132 km from North to South.

The County administrative headquarters is at Kapenguria town and the County is divided into

four districts, 14 divisions, 58 locations and 188 sub-locations.

In times of peace between the Pokots who are the main inhabitants of West Pokot County and

the Karamojong of Uganda, livestock (camels included) graze and browse across the border into

Uganda.

24

Source: Ministry for development of Northern Kenya and other arid lands

Figure 3.2: Map of West Pokot County showing the six study divisions

Rainfall is bimodal with the long rains falling between March and June and the short rains

occurring from September to November. The rainfall amounts range from 700 mm in the

lowlands to 1600mm in the high altitude zones. Temperatures in the lowlands range from15ºC to

30ºC but the highlands may experience temperatures as low as 9ºC. The major drainage systems

in the County are Turkwel, Kerio and Nzoia rivers. Both the Turkwel and Kerio Rivers drain

Northwards into Lake Turkana while Nzoia River drains into the Lake Victoria in the South.

25

Traditionally, the Pokots are nomadic pastoralists whose lifestyle is rapidly changing to

sedentary mixed farmers, especially in areas where conditions permit. Like many other arid and

semi arid areas in the country, the area has been experiencing rapid population growth for both

human and livestock. Physical infrastructure such as roads, telecommunications, hospitals and

schools are poorly developed. The harsh climatic conditions over most of the area and difficult

terrain make a large proportion of the County inaccessible. Traditional mobile pastoral lifestyle

is practiced by most of the community members. The sustainable utilization of natural resources

in the County is hampered by mainly socio-economic and technical capacity.

Insecurity was rampant in the County in the past and often involved theft of livestock, but

there has been major shift towards the restoration of peace with neighbouring communities.

Conflict over grazing resources is also common in part due to the breakdown of traditional

pasture management systems and increasing individual herds. Insecurity especially across the

border makes sustainable utilization of livestock resources difficult.

Livestock is the most important economic resource in the County and supports the main

livelihood system. The main livestock species found in this County in order of importance are

cattle, goats, sheep, donkeys, camels and poultry. There is, however, a gradual change in this

order in response to population pressure, competition and availability for pasture. Consequently,

agro-pastoralism is taking root in the County with a marked increase in crop farming where

possible.

Another discernible shift is the rising popularity of goats and camels in relation to cattle

in view of limited grass availability and more browse availability. Emerging livestock, especially

poultry is also rising in popularity as women and youth who are not traditionally allowed to own

cattle keep them and can easily sell them to raise money in periods of emergency need.

26

There are three sub counties which make up West Pokot County namely Pokot North, Pokot

Central and West Pokot sub counties. The estimated livestock population is shown below in

Table 3.1.

Table 3.1: Estimated livestock population in West Pokot County

Sub county Cattle Sheep Goats Camels Donkeys

West Pokot 129,273 114,050 173,693 294 8,243

North Pokot 377,688 199,977 377,903 29,273 21,671

Pokot Central 179,212 146,300 213,141 1,050 6,559

Total 686,375 460,327 764,737 30,617 36,473

Source: Kenya National Bureau of Statistics, National Census 2009

3.1.2 Study Design

A cross sectional study was undertaken on 95 lactating and traditionally managed one-

hump camels (Camelus dromedarius) in several selected households (manyattas). These

manyattas were the sampling units. The study took place during the months of August and

November 2012 in Kongelai, Kacheliba, Konyao, Kasei, Kiwawa and Alale divisions of West

Pokot County (Figure 3.2).These divisions are the only divisions with the highest population of

camels in West Pokot County and they are located along the border between Kenya and Uganda.

27

3.1.3 Selection of the study sites, households and camels

The divisions within West Pokot County were selected based on the sizeable population

of camels, good security and passable roads. The local District Veterinary Officer (DVO) & the

District Livestock Production Officer (DLPO) were engaged in the sensitization and

mobilization of the camel producers so that the producers would be aware of this study and to

explain their expected role in the study. The Kenya Camel Association (KCA) which is an

umbrella Association for camel producers in Kenya was also involved in the mobilization of the

camel keepers through the local KCA Regional Representatives in the County. The DVOs,

DLPOs and KCA prepared the list frame of all the eligible camel owners in each study division

within the county. In addition, they indicated the manyattas where there was at least one lactating

camel.

All the eligible sampling units along the main road (transect two km on either side) from

the South of the County to the furthest division in the North were selected through a systematic

random sampling method. The number of camels sampled in each division was proportional to

the population of camels in the division. The process continued until the required sample size

was reached.

3.1.4 Data collection

Data on household demographics (human and animal) and known and/or reported

diseases of camels, and animal level factors (each camel) such as age, parity number, stage of

lactation, breed, current milk yield, milk abnormalities, whether the camel has had mastitis

before and presence of udder/teat lesions were collected and recorded in a semi-structured

28

questionnaire (SSQ) (Appendix I) during sampling. Data were gathered through interviews

administered to the household head or any other household member conversant with the camel

management.

The age of the camels was estimated (by observing the eruption and wearing of the front

permanent teeth) since there were no records available and were categorized as young adults

(>4yrs to 6yrs), adults (≥ 6yrs to ≤ 8yrs) and old >8yrs. The stage (length) of lactation was

categorized as early (1st to 4th month), mid (> 4th month to 8th month) and late (> 8th month).

Number of parity was categorized as few (≤ 3 calves), moderate (4 -7 calves) and many (> 7

calves).

The SSQ were also used to capture data from direct observations in addition to one-on-one

interviews with the camel owners.

Data collected at herd-level included; herd size, the producer’s knowledge on treatment attempts

& control of mastitis (both conventional and traditional methods), milking frequencies &

procedures such as washing the udder with clean warm water before milking.

3.1.5 Sample size determination for the number of milk samples

The following formula was used to calculate the sample size (Dohoo et al., 2003):

n = Zα2pq/L2,

where, n=sample size, Zα = the value of z that gives 95% confidence interval

(1.96), p = a priori prevalence (estimated prevalence), q = 1-p and L = Allowable error.

The prevalence of mastitis in camels in Kenya was estimated at 25% as was reported by Younan

et al (2001). Thus adopting a p of 25% and L of 5%, then;

n = (1.962*0.25*0.75)/ (0.05)2 = 290 (quarter milk samples).

29

3.1.6 Collection of milk samples

Prior to the commencement of sample taking the camel owner’s consent was obtained

after explaining the purpose of the study. Milk samples were then aseptically collected from

each individual quarter from 95 lactating camels (14 from Kongelai, 10 from Kacheliba, 2 from

Konyao, 37 from Kiwawa, 23 from Kasei and 9 from Alale division) during either morning,

midday or evening milking time depending on what was logistically convenient. This was done

after the calf was allowed to suckle to allow milk let down. Visual examination (by observation

and palpation) of all the quarters was also carried out.

About 20 ml of milk was collected from each quarter directly into clean and sterile

sample bottles that were clearly labeled and immediately stored in cool boxes with ice packs

before refrigeration in the evening at around 50C and later transported in cool boxes with ice

packs to the Bacteriology Laboratory at the Department of Veterinary Pathology, Microbiology

and Parasitology, Faculty of Veterinary Medicine, University of Nairobi for bacterial culture.

3.1.7 Examination for sub-clinical mastitis at household level

This was done by carrying out California Mastitis Test (CMT) using the method

described by Schalm and Noorlander (1957) immediately after sample collection an equal

volume (about 2ml) of CMT reagents (Delaval, Poland) mix with an equal volume (about 2ml)

of sampled milk in each segment of the CMT paddle and mixed gently. The results were read

and visually scored for each quarter within ten seconds depending upon the amount of gel

formation as follows:

30

Negative = no reaction.

Trace = appearance of streaks that can be made visible during rotation of the plate.

1+= distinct thickening during rotation, but no gel.

2+ = slight formation of gel which follows the rotating plate very slowly.

3+ = solid formation of gel that adheres to the base of the plate.

Quarters whose scores were negative, trace and 1+ were considered healthy while scores ≥ 2+

were considered infected or positive for subclinical mastitis. The test mixture (milk sample and

the CMT reagent) was discarded and the paddle washed with clean water after each use to enable

it to be used in the next selected lactating camel.

3.1.8 Bacteriological Examination and Isolations

After culturing, bacteriological examination was carried out following standard methods

laboratory and field handbook on bovine mastitis – (1987), Sears et al (1993) and Quinn et al

(1994) to identify major bacterial agents associated with mastitis. In brief, milk samples from the

deep freezer were thawed to room temperature and one loopful (10ul or 0.01ml) of the samples

was aseptically streaked on Blood Agar (5% defibrinated sheep blood) plates and MacConkey

Agar (MA) plates (Carter et al, 1991). Bacterial growths were identified and recorded after

incubation for 24 to 48 hours at 37ºC aerobically. Primary cultures were considered to be

positive when bacterial growth was observed on the inoculated plates and negative when no

bacterial growth was observed.

Pure culture was further obtained by sub-culturing part of typical and well isolated

colony on a corresponding medium and incubated further at 37ºC aerobically for 24 hours.

31

Identification of bacterial isolates was done based on colony morphological features and

hemolytic reactions (primary cultures), gram staining reactions and biochemical tests on pure

cultures (Quinn et al., 1994). Gram stain procedures were performed according to the method

described by (Quinn et al., 1998; Bebora et al., 2007 and Forbes et al., 2007). To differentiate

Staphylococcus and Streptococcus spp, catalase reaction was performed on all Gram- positive

isolates employing the rapid slide technique described by Cheesburgh (2000). A drop of 3%

hydrogen peroxide was placed on a slide, organism was added & mixed and observed for

bubbling to confirm the presence of catalase enzyme. Catalase negative reaction indicated

presence of Streptococcus spp whereas catalase positive indicated Staphylococcus spp.

Coagulase test was carried out to differentiate Staphylococcus aureus from other Staphylococcus

spp. CAMP (Christie, Atkins and Munch-Petersen) test and growth in MacConkey agar plate

was also carried out to differentiate Streptococcus agalactiae from other mastitis causing

Streptococcus.

3.1.9 Statistical Data Analysis

All data collected were entered in Microsoft Excel 2007 worksheet as database and

exported to Instat Plus for statistical analysis. Descriptive statistics were generated using the

same statistical package. Differences in proportions were assessed using the chi square at 5%

level of significance in univariate analysis.

The odds ratio (OR) was used to assess the strength of any associations identified,

initially in the logistic regression univariate analysis to screen variables (p<0.1) and later

multivariate logistic regression models were used to test the above variables for significance

32

(p<0.05). Significance of risk factors on the presence of mastitis (the variable outcome) was

calculated using chi-square (x2) technique to test the existence of statistical association between

mastitis and the risk factors (explanatory variables) such as age, parity, stage of lactation and

breed. In all chi-square test applications level of p<0.05 was considered statistically significant.

In addition logistic regression analysis was used to calculate the odds ratio (OR) to measure the

degree/strength of association between the risk factors and the presence of mastitis in camel.

3.1.10 Definition of outcome variables

a) Subclinical mastitis defined by CMT

All CMT results/scores were judged as follows; trace and 1+ considered negative and 2+, 3+, 4+

& 5+ considered positive results. A quarter was considered CMT-Positive if it had a score of ≥

2+ while a camel was defined CMT-Positive if it had at least one quarter with a CMT score ≥ 2+.

b) Subclinical mastitis defined by microbiological cultures

A quarter was defined as positive when a pathogen (bacterium) was isolated during

microbiological culture. A camel was considered positive when at least one quarter milk sample

had a pathogen isolated. When a herd had a camel with mastitis, that herd was considered

positive for mastitis.

33

CHAPTER FOUR

4.1 RESULTS

4.1.1 Characterization of camel management practices

The results of the study showed that, many Pokot camel owners generally milked their

camels thrice a day (90.4% (47/52)); at dawn or early in the morning; at 10 am (limo in Pokot)

and at night about 8.00pm (2-3 hours after returning from grazing). Only 9.6% (5/52) milked

their camels twice a day. Majority of the milkers (82.7% (43/52)) washed their hands once

before milking all the livestock beginning with goats, cows and lastly the camels; while 17.3%

(9/52) did not wash their hands at all. On washing the udder before milking, 76.9% (40/52) did

not wash the udder. They believed that the calf would clean the udder by suckling. About 7.7%

(4/52) washed the udder before milking. These were mostly camel keepers around trading

centres. Those who did not respond to the practice of washing udder before milking were 15.4%

(8/52) (Table 4.1).

Most (75% (39/52)) of the treatment of camel mastitis was done by the owners. Few

(26.9% (14/52)) went for some assistance from the Community-based animal health workers

(CBAHWs). Veterinarians (VOs) and animal health assistants (AHAs) were not consulted on

treatment of mastitis. During mastitis infection, 75% (39/52) did not follow the milking order by

starting to milk the clean camels first and the infected camels last. The rest 25% (13/52) (as

shown in Table 4.1) abandoned milking the infected camel and left the calf to suckle. They

claimed the camel was feeling pain during milking.

34

The camels were herded during the daytime on communal grazing lands and kept at night in

traditional enclosures (bigh in Pokot) made of thorny bushes and tree branches as protection

from predators. At night the calves were penned separately but during the day they accompanied

their mothers and suckled freely. The milking frequency reduced or increased depending on the

season, yield and the stage of lactation of the camels. It was also observed that the Pokots did not

milk their camels outside the homesteads.

Table 4.1: Camel management practices

Household practices Frequency Percentage (%)

Milking Rates Thrice a day 47 90.4 Twice a day & below 5 9.6

Wash hands before milking

Yes 43 82.7 No 9 17.3

Wash the udder before milking

Yes 4 7.7 No 40 76.9 No respond 8 15.4

Who milks the camels Owners 14 26.9 Wives 16 30.8 Herders (young boys) 22 42.3

Who treats camels with mastitis

V.O 0 0 AHA 0 0 Self 39 75 CBAHWs 14 26.9

Follow milking order during mastitis

Yes 13 25 No 39 75

It was also observed during the study that the Pokot herders traditionally isolated near term she-

camels and even after calving from the rest of the herd during the day and night for a month

35

before the calf was allowed to accompany the mother during the day for grazing. If a she-camel

calved far away from the homestead she was left to remain with the calf for about three days

before driven back to the homestead.

Amongst the Pokot people, camels were mostly milked by young boys (herders) (42.3%

(22/52)) and adult women (30.8% (16/52)). In the absence of women, the men (26.9% (14/52))

could also milk the camels. During milking they allowed the calf to suckle to stimulate milk let

down. It was also observed that the Pokot pastoralists kept camels together with other livestocks

such as cattle, sheep and goats in the same Manyatta.

The herders also reported that camel mastitis existed in West Pokot and they believed it affected

other livestock and even human beings (women). Majority of the respondents (80.8% (42/52)) as

shown in Table 4.2 reported that they had traditional ways of managing the infection which

included the following; treating using local herbs 26.9% (14/52) (using leaves, roots & exudates

from various plants), 11.5% (6/52) milking the affected udder into a red hot Panga so that the

smoke goes back to the udder to treat it, 21.2% (11/52) branding with hot Iron, 17.3% (9/52)

smear with accaricide (dip) to prevent flies especially in the case of gangrenous mastitis. Two

pastoralists (3.8%) from Kasei division reported they used dawa nyoka (anti-venom) to treat

mastitis because they associated camel mastitis with snake bites.

36

Table 4. 2: Traditional ways of managing camel mastitis in West Pokot County

Traditional remedies Frequency Percentage (%) Using local herbs (leaves, roots) 14 26.9 Using hot Iron 11 21.2 Using accaricide (dip) 9 17.3 Using hot Panga 6 11.5 Using dawa nyoka (anti-venom) 2 3.8 Total 42 80.8

The study further showed that the most commonly reported camel diseases by the camel keepers

included Mange, Trypanosomiasis, mastitis, Haemorragic Septiceamia (HS) and abscesses as

shown in Table 4.3. The results showed that mastitis was mentioned among the important

diseases.

Table 4. 3: Camel diseases and conditions reported by Pokot herders

Disease or Condition Local (Pokot) name Frequencies % Mange Simbirion 36 69.2 Trypanosomiasis Plis 34 65.4 Mastitis Semewo Krusho 29 55.8 Haemorrgic Septiceamia (HS) Chemotow 26 50.0 Abscesses Pirieng’wa 15 28.8 Wounds Moyoi 11 21.2 Diarrhoea Kiyitagh 9 17.3 Pneumonia Psosoi 9 17.3 Tick infestation (especially in camel calves)

Tilis 8 15.4

Camel Orf Ng’rumen 8 15.4 Sudden death Lotuler 5 9.6 Abortion Toronogh 3 5.8 Worms (Helminthiasis) Chepturu 3 5.8

37

4.1.2 Household demographics

4.1.2.1 Age and sex distribution

The 52 households interviewed during the study were distributed as follows: Kongelai division

(n=5), Kacheliba division (n=7), Konyao division (n=2), Kiwawa division (n=14), Kasei division

(n=20) and Alale division (n=4).

Of the 52 households interviewed 86.5% (45/52 households) were headed by males and 13.5%

(7/52 households) were headed by females. 100% (52/52) of the households were settled in

communal land; therefore most of the camels were reared in communal range land. The age

brackets of the camel owners were <20 yrs 3.8% (2/52); 21-30 yrs 15.4% (8/52); 31-40 yrs

36.5% (19/52); 41-50 yrs 26.9% (14/52); 51-60 yrs 9.6% (5/52) and >60 yrs 7.7% (4/52) (Table

4.4 & Figure 4.1). The majority of the camel owners were therefore between 31 & 40 years of

age. The least owners of camels were below 20 yrs and above 60 yrs of age.

Table 4. 4: Age and sex distribution of the household's heads interviewed

Age (yrs) <20 21-30 31-40 41-50 51-60 >60 Total

Male 2 5 18 12 4 4 45 Female 0 3 1 2 1 0 7 Total 2 8 19 14 5 4 52

38

Figure 4.1: Age and sex distribution of the households' heads interviewed

4.1.2.2 Herd structure

The herd structure in the study area showed that there were more (42.9% (67/165))

lactating camels in Kiwawa division compared to other divisions. The least (2.6% (4/156))

number of lactating camels were in Konyao division. There were also more dry herds (41.1%

(109/265)) in Kiwawa division than any other division. In overall, there were more (37.4%

(326/872) camels in Kiwawa division in comparison to other divisions. Samples were collected

from 95 lactating camels from the total of 165 lactating camels in 52 herds in the study areas.The

herd size range from 3 to 50 camels with mean herd size of 16.8±. Forty four herds (84.6%

(44/52)) tested positive for sub-clinical mastitis against 8 herds (15.4% (8/52)) which tested

negative. The rest are as shown in Table 4.5.

0

2

4

6

8

10

12

14

16

18

20

< 20 21-30 31-40 41-50 51-60 61-70

male

Female

39

Table 4.5: Summary of herd structure per division

Division Lactating camels

Dry camels

Breeding bulls

Immature males

Immature females

Female calves

Male calves

Total

Kongelai 22 20 7 17 10 9 12 97 Kacheliba 17 27 6 16 26 12 7 110 Konyao 4 13 1 6 6 2 2 34

Kasei 36 51 4 30 31 16 19 187 Kiwawa 67 109 8 32 46 29 36 326 Alale 19 45 3 13 18 13 7 118 Total 165 265 29 114 137 81 99 872

4.1.3 Laboratory Results on bacterial culture and identification

A total of 380 quarter milk samples (56 from Kongelai division, 40 from Kacheliba

division, 8 from Konyao division, 148 from Kiwawa division, 92 from Kasei and 36 from Alale)

were collected during the study. The same samples from 95 apparently clinically healthy

dromedary camels were cultured in the laboratory to identify subclinical mastitis and its

causative agents.

4.1.3.1 Prevalence of subclinical mastitis at animal (camel) level in the study area

The results have shown that the prevalence of subclinical mastitis at animal level was highest in

Kongelai division (100% (14/14)) followed by Kasei division (82.6 % (19/23)) then Kacheliba

division (80% (8/10)). The lowest prevalence was in Konyao division at 0% (0/2) prevalence as

shown in Table 4.6.

40

The overall animal (camel)-level prevalence of subclinical mastitis in the study area was 76.8%

(73/95) out of which 23.2% (22/95 camels) had only one mammary quarter affected, 21.1%

(20/95 camels) had two quarters affected, 17.9% (17/95 camels) had three quarters affected

while 14.7% (14/95camels) had all the four quarters affected.

Table 4.6: Prevalence of subclinical mastitis at animal (camel) level in the study area

Division Positive Negative Total Prevalence (%)

Kongelai 14 0 14 100 Kasei 19 4 23 82.6 Kacheliba 8 2 10 80 Kiwawa 27 10 37 73 Alale 5 4 9 55.6 Konyao 0 2 2 0 Total 73 22 95 76.8

4.1.3.2 Quarter infection rates

Out of the 380 quarter samples cultured for bacteria, 44.5% (169/380) tested positive for

subclinical mastitis giving a prevalence of quarter-level mastitis at 44.5% while the rest 55.5%

(211/380) of the samples tested negative (Table 4.7 & Appendix II).

Table 4.7: Prevalence of subclinical mastitis at quarter level

Quarter Quarter Positive Negative Total Prevalence (%)

Right Fore quarter 43 52 95 11.3 Right Hind quarter 46 49 95 12.1 Total 89 101 190 Left Fore quarter 40 55 95 10.5 Left Hind quarter 40 55 95 10.5 Total 80 110 190

41

The results also showed that the right quarters were more affected compared to the left quarters

(12.1% (46/380) prevalence for RHQ and 11.3% for RFQ vs 10.5% each in both LFQ & LRQ)

(Appendix II).

The Mhor & MHCh-2 (Mantel-Haenszel Chi-square) were 0.94 & 0.17 respectively while the

adjusted (summary) odds ratio (OR) was 0.88. This showed that mastitis infection and the

quarters of the camel udder were not significantly associated.

The results further showed that the RHQ was the most frequently infected quarter at a prevalence

of 27.2% (46/169) followed by the RFQ at a prevalence of 25.4% (43/169) (Figure 4.2). The two

left quarters (LFQ &LHQ) were least infected as shown in Figure 4.2.

Figure 4.2: Prevalence of subclinical mastitis per quarter

Table 4.7 above was collapsed into two 2x2 tables as shown below in Tables 4.8 & 4.9.

21

22

23

24

25

26

27

28

RFQ RHQ LFQ LHQ

Prevalence (%)

42

Table 4.8: Prevalence of subclinical mastitis at two quarters (left & right) level

Quarters Positive Negative Total Prevalence (%)

Odds Ratio (OR)

Chi-square P- value

Right quarters 89 101 190 46.8 1.2 0.86 0.05 Left quarters 80 110 190 42.1 Total 169 211 380

Table 4.9: Prevalence of subclinical mastitis at quarter level showing OR & Chi-2

Quarters Positive Negative Total Prevalence

(%) Odds Ratio (OR) Chi-2 P- value

Front Quarters 83 107 190 43.7 0.94 0.1 0.05 Hind Quarters 86 104 190 45.3 Total 169 211 380

The results from the two tables above showed that there were no associations between the

position of the quarters and the occurrence of mastitis in camels at p>0.05. This is because the

calculated chi-square values in both tables are less than the critical value of 3.84 at 95%

confidence.

4.1.4 Association between the occurrence of mastitis and other risk factors

Among many potential explanatory variables, two were considered as potential risk

factors for the occurrence of sub clinical mastitis in this study. These were stage of lactation and

parity of the lactating camels. The association of subclinical mastitis with these risk factors using

chi-square and odds ratio (OR) are as shown in Table 4.10 and 4.11. The calculated values of

both Chi-square and OR in Tables 4.9 & 4.10 showed that there was a significant association

between the two risk factors (stage of lactation and parity) and subclinical mastitis at P<0.05.

43

This is because the calculated values of Chi-square (4.08 & 4.48 in both tables) were greater than

the critical value of 3.84. The risk of subclinical mastitis infection in camels more than two

months of lactation was 2.75 higher than in camels in less than two months in lactation.

Table 4.10: Association between the occurrence of mastitis and stage of lactation

Lactation Stage

Positive Negative Total Prevalence (%)

OR Chi-2 P- value

old (> 2 month)

56 12 68 82.4 2.75 4.08 0.05

young (≤ 2

month) 17 10 27 63.0

Total 73 22 95

The results also showed that lactating camels with parity of more than two calvings were 2.9

times more likely to be infected by mastitis than camels of lower or equal to two calving parity

as shown in Table 4.11.

Table 4.11: Association between the occurrence of mastitis and parity

Parity Positive Negative Total Prevalence (%) OR Chi-2 p-value > 2 calving 42 7 49 85.7 2.90 4.48 0.05 ≤ 2 calving 31 15 46 67.4 Total 73 22 95

44

4.1.5 Bacterial Isolation Analysis

The bacteria isolated from the 380 quarter samples are shown in Table 4.12. A total of

114 bacteria were isolated with the most predominant bacterium being Staphylococcus aureus

with prevalence of 36.0% (49/136), followed by E. coli with prevalence of 27.2% (37/136).

Streptococcus agalactiae & Staphylococcus epidermidis were the third predominant isolates with

prevalence of 9.6% (13/136) each. Micrococcus spp & Pseudomonas were the least isolates with

less than 1% prevalence. A diagnosis of ‘no bacterial growth’ was made in 22 cases which is

16.2% (22/136). There were no contaminated samples recorded. Overall all milk samples

produced mixed types of bacterial growth in the primary cultures. This indicated that there was a

multiple infection of the quarters.

Identical pathogens were also isolated from different quarters of individual camels and from

camels within the same herd suggesting that transmission from quarter to quarter and camel to

camel had occurred.

Table 4.12: Bacterial isolation rates and their prevalences

Micro-organism Total number of Isolates (n) Prevalence (%)

Nil (no growth) 22 16.2

Staphylococcus aureus 49 36.0

Streptococcus agalactiae 13 9.6 Escherichia coli 37 27.2

Staphylococcus epidermidis 13 9.6

Micrococcus species 1 0.7

Pseudomonas 1 0.7

Contaminated 0 0

Total 136 100

45

4.1.5.1 Bacterial Isolation per division

The results showed that there were more bacterial isolations in Kiwawa division (36% (41/114))

followed by Kasei division at 22.8% (26/114). There was no isolation of pathogens in Konyao

division. The highest isolation of Staphylococcus aureus (36.7% (18/49)) was in Kiwawa

division while Alale division had the lowest isolation at 8.2% (4/49). Escherichia coli isolation

was also high (32.4% (12/37)) in Kiwawa division followed by Kasei division at 24.3% (9/37) as

shown in Table 4.13.

Table 4.13: Bacterial Isolation per division

Division S. aureus E. coli S. epidermidis S. agalactiae Micrococci spp

Pseudomonas Total Prevalence (%)

Kongelai 7 7 5 5 - - 24 21.1 Kacheliba 7 6 - - - - 13 11.4 Konyao - - - - - - - 0 Kiwawa 18 12 3 8 - - 41 36 Kasei 13 9 2 - 1 1 26 22.8 Alale 4 3 3 - - - 10 8.8 Total 49 37 13 13 1 1 114

46

4.2 Results per Division

4.2.1 Kongelai Division

4.2.1.1 Herd Structure by age of camel in Kongelai division

Lactating camels were 22.7% (22/97) which gave an average of 4.4 (22/5) lactating camels per

household. Of these 63.6% (14/22) were sampled. The dry herd and the breeding bulls were

20.6% (20/97) and 7.2% (7/97) respectively as shown in Table 4.14 and Figure 4.3 below. Most

(100%) of the camels were Turkana breeds.

Table 4.14: Herd structure by age of camel in Kongelai division

Household (HH)

Lactating camels

Dry camels

Breeding bulls

Immature males

Immature females

Female calves

Male calves

Total

1 10 8 3 6 5 4 6 42 2 4 3 1 5 2 1 2 18 3 3 2 1 2 1 1 2 12 4 3 4 1 1 0 2 1 12 5 2 3 1 3 2 1 1 13 Total 22 20 7 17 10 9 12 97

47

Figure 4.3: Herd structure in Kongelai division

4.2.1.2 Prevalence of subclinical mastitis at animal (camel) level in Kongelai division

The results of the bacterial culture from the 14 camels indicated that at least one quarter

milk sample had a pathogen isolated from each camel. Therefore at camel (animal)-level the

prevalence of subclinical mastitis in Kongelai division was 100% (14/14). About 14.3% (2/14

camels) had only one quarter infected, 42.9% (6/14 camels) had two quarters infected, 21.4%

(3/14 camels) had three quarters infected while 21.4% (3/14camels) had all the four quarters

infected) with Staphylococcus aureus being the predominant bacterium isolated from the milk

samples.

Lactating camels23%

Dry camels21%

Breeding bulls7%

Immature males18%

Immature females

10%

Female calves9%

Male calves12%

Herd structure Kongelai division

48

4.2.1.3 Prevalence of subclinical mastitis at quarter level in Kongelai division

Of the 56 quarter samples cultured from Kongelai division 64.3% (36/56) quarter milk

samples were positive for subclinical mastitis while 35.7% (20/56) were negative. The right fore

quarter was the most affected 92.9% (13/14) and the right hind quarter was the least affected

35.7% (5/14). The overall quarter-level prevalence of subclinical mastitis in this division was

64.3% (36/56).

Table 4.15: Prevalence of subclinical mastitis at quarter level in Kongelai division

Quarter Positive Negative Total Prevalence (%) RFQ 13 1 14 92.9 RHQ 5 9 14 35.7 LFQ 8 6 14 57.1 LHQ 10 4 14 71.4 Total 36 20 56

49

4.2.2 Kacheliba Division

4.2.2.1 Herd structure by age of camel in Kacheliba division

Samples were collected from 10 lactating camels from the total 17 lactating camels in seven

households selected in Kacheliba division. All (100%) the camels in the division were of

Turkana breed. The lactating herd was 15% (17/110) which gave an average of 2.4 (17/7)

lactating camels per household in the division. The dry herd and the breeding bulls were 24%

(27/110) and 5% (6/110) respectively.

Table 4.16: Herd structure by age of camel in Kacheliba division

HH No

Lactating camels

Dry camels

Breeding bulls

Immature males

Immature females

Female calves

Male calves

Total

1 3 2 1 3 3 3 0 15

2 2 2 1 1 5 2 0 13 3 2 4 1 0 3 1 3 14 4 2 2 0 2 0 1 1 8 5 2 2 0 1 4 1 1 11 6 2 8 2 4 5 1 1 23 7 4 7 0 5 6 3 1 26

Total 17 27 6 16 26 12 7 110

50

Figure 4. 4: Herd structure Kacheliba division of West Pokot County

4.2.2.2 Prevalence of subclinical mastitis at animal (camel) level in Kacheliba division

At the animal (camel)-level, the prevalence of mastitis was 80% (8/10). About 30%

(3/10) had only one quarter affected, 30% (3/10) had two quarters affected, 10% (1/10) had three

quarters affected while 10% (1/10) had all the four quarters infected. This was 20% lower than

Kongelai division. Staphylococcus aureus and E. coli were the major causative agents of camel

subclinical mastitis in Kacheliba division.

Lactating camels15%

Dry camels24%

Breeding bulls5%

Immature males15%

Immature females

24%

Female calves11%

Male calves6%

Herd structure in Kacheliba division

51

4.2.2.3 Prevalence of subclinical mastitis at quarter level in Kacheliba division

In this division, there were 40 quarter samples cultured out of which 40% (16/40) quarter

milk samples were positive for subclinical mastitis while 60% (24/40) were negative, therefore at

quarter level the prevalence of subclinical mastitis was 40% (16/40). The left hind quarter was

the most affected 60% (6/10) followed by the right hind quarter 40% (4/10). The fore quarters

were least infected 30% (3/10) each.

Table 4. 17: Prevalence of subclinical mastitis at quarter level in Kacheliba division

Quarter Positive Negative Total Prevalence (%) RFQ 3 7 10 30 RHQ 4 6 10 40 LFQ 3 7 10 30 LHQ 6 4 10 60 Total 16 24 40

52

4.2.3 Konyao Division

4.2.3.1 Herd structure by age of camels in Konyao division

Two households were sampled in this division and just like other divisions most camels were of

Turkana breeds. Lactating camels were 12% (4/34) and 100% (4/4) were sampled.

Table 4.18: Herd structure by age of camels in Konyao division

Household (HH)

Lactating camels

Dry camels Breeding bulls

Immature males

Immature females

Female calves

Male calves

Total

1 2 3 0 0 2 2 0 9 2 2 10 1 6 4 0 2 25 Total 4 13 1 6 6 2 2 34

Figure 4.5: Herd structure Konyao division of West Pokot County

Lactating camels12%

Dry camels38%

Breeding bulls3%

Immature males17%

Immature females

18%

Female calves6%

Male calves6%

Herd structure Konyao division

53

4.2.3.2 Prevalence of subclinical mastitis at animal (camel) level in Konyao division

There was no single pathogen isolated from the quarter milk samples taken from camels in the

division, therefore at camel (animal) level the prevalence of subclinical mastitis was 0%. From

the results, the two sampled herds were clean.

4.2.3.3 Prevalence of subclinical mastitis at quarter level in Konyao division

There were no pathogens isolated from the eight quarter milk samples taken from the division.

This gave 0% (0/8) prevalence of subclinical mastitis at quarter level.

Table 4. 19: Prevalence of subclinical mastitis at quarter level in Konyao division

Quarter Positive Negative Total Prevalence (%) RFQ 0 2 2 0 RHQ 0 2 2 0 LFQ 0 2 2 0 LHQ 0 2 2 0 Total 0 8 8

54

4.2.4 Kiwawa Division

4.2.4.1 Herd structure by age of camel in Kiwawa division

Samples were taken from 37 lactating camels from the total 67 lactating camels in the 14

households selected in the division. The lactating camels were 20.6% (67/326) which gave an

average of 4.8 (67/14) lactating camels per household in the division. Samples were taken from

52.2% (37/67) of the lactating camels as shown in Table 4.20 and Figure 4.6 below.

Table 4.20: Herd structure Kiwawa division

Household (HH)

Lactating camels

Dry camels

Breeding bulls

Immature males

Immature females

Female calves

Male calves

Total

1 5 9 1 2 2 3 2 24 2 7 30 2 0 3 3 4 49 3 2 18 1 7 6 1 1 36 4 4 5 0 1 4 3 1 18 5 4 5 0 1 5 1 3 19 6 4 5 1 3 4 1 3 21 7 3 2 0 2 2 1 2 12 8 8 26 1 2 3 3 5 48 9 3 2 0 0 0 1 1 7 10 2 0 1 5 2 1 1 12 11 2 0 0 0 0 0 2 4 12 6 0 0 0 0 2 4 12 13 10 5 1 2 8 7 3 36 14 7 2 0 7 6 2 4 28 Total 67 109 8 32 46 29 36 326

55

Figure 4.6: Herd structure in Kiwawa division of West Pokot County

4.2.4.2 Prevalence of subclinical mastitis at animal (camel) level in Kiwawa division

At the animal (camel) level the prevalence of mastitis was 73% (27/37). Majority of the

animals 27.0% (10/37) had only one quarter affected, 16.2% (6/37) had two quarters affected,

also 16.2% (6/37) had three quarters affected and 13.5% (5/37) had all the four quarters infected.

Staphylococcus aureus and E. coli were the major causative agents of camel subclinical mastitis

in the division.

4.2.4.3 Prevalence of subclinical mastitis at quarter level in Kiwawa division

At quarter level the prevalence of subclinical mastitis in the division was 40.5% (60/148). The

right hind quarter was the most affected 48.6% (18/37) followed by the right front quarter at

40.5% (15/37). The left quarters were the least infected.

Lactating camels21%

Dry camels33%

Breeding bulls2%

Immature males10%

Immature females

14%

Female calves9%

Male calves11%

Herd structure in Kiwawa division

56

Table 4. 21: Prevalence of subclinical mastitis at quarter level in Kiwawa division

Quarter Positive Negative Total Prevalence (%) RFQ 15 22 37 40.5 RHQ 18 19 37 48.6 LFQ 14 23 37 37.8 LHQ 13 24 37 35.1 Total 60 88 148

57

4.2.5 Kasei Division

4.2.5.1 Herd structure in Kasei division

The lactating camels were 19.3% (36/187) which gave an average of 1.8 (36/20) lactating camel

per household. The dry camels were 27.3% (51/187) while the breeding bulls were 2.1% (4/187).

Samples were taken from 63.9% (23/36) of the lactating camels in 20 households selected in the

division.

Table 4.22: Herd structure in Kasei division

Household (HH)

Lactating camels

Dry camels

Breeding bulls

Immature males

Immature females

Female calves

Male calves

Total

1 1 3 0 0 0 0 1 5 2 2 2 0 1 0 1 1 7 3 4 5 0 3 4 1 3 20 4 1 1 0 0 0 1 0 3 5 2 3 0 2 1 0 2 10 6 2 5 0 3 3 2 0 15 7 3 5 2 3 2 2 1 18 8 2 1 0 0 0 0 2 5 9 1 1 0 0 1 0 1 4 10 1 4 1 0 2 1 0 9 11 5 3 1 10 4 3 2 28 12 1 2 0 1 3 0 0 7 13 1 3 0 0 3 1 0 8 14 2 2 0 2 3 1 1 11 15 1 3 0 1 0 0 1 6 16 1 2 0 1 0 0 1 5 17 1 1 0 0 1 0 1 4 18 2 3 0 3 2 1 1 12 19 2 0 0 0 2 1 1 6 20 1 2 0 0 0 1 0 4 Total 36 51 4 30 31 16 19 187

58

Figure 4.7: Herd structure in Kasei division of West Pokot County

4.2.5.2 Prevalence of subclinical mastitis at animal (camel) level Kasei division

From the results obtained the prevalence of subclinical mastitis at animal (camel) level was

82.6% (19/23). In terms of quarters affected 21.7% (5/23) had only one quarter affected, another

21.7% (5/23) had two quarters affected, also another 21.7% (5/23) had three quarters affected

and 17.4% (4/23) had all the four quarters affected. Most of the udder infections were mixed and

caused by both Staphylococcus aureus and E. coli.

Lactating camels19%

Dry camels27%

Breeding bulls2%

Immature males16%

Immature females

17%

Female calves9%

Male calves10%

Herd structure Kasei division

59

4.2.5.3 Prevalence of subclinical mastitis at quarter level Kasei division

The overall quarter-level prevalence of subclinical mastitis in the division was 53.3% (49/92).

The most affected quarter was the right hind-quarter 65.2% (15/23) followed by the right front

quarter 56.5% (13/23) while the least affected was the left hind-quarter 39.1% (9/23).

Table 4.23: Prevalence of subclinical mastitis at quarter level in Kasei division

Quarter Positive Negative Total Prevalence (%) RFQ 13 10 23 56.5 RHQ 15 8 23 65.2 LFQ 12 11 23 52.2 LHQ 9 14 23 39.1 Total 49 43 92

60

Alale Division

4.2.6.1 Herd structure Alale division

Lactating camels were 16.1% (19/118) which gave an average of 4.75 lactating camels per

household. Samples were taken from 47.4% (9/19) of the lactating camels. The dry herd was

38.1% (45/118) while the bulls were 3% (3/118) of the total herds in the division as shown in

Table 4.24 and Figure 4.8.

Table 4.24: Herd structure Alale division

Household (HH)

Lactating camels

Dry camels

Breeding bulls

Immature males

Immature females

Female calves

Male calves

Total

1 6 30 1 7 0 5 1 50 2 4 1 0 0 0 2 2 9 3 5 14 1 1 4 4 2 31 4 4 0 1 5 14 2 2 28 Total 19 45 3 13 18 13 7 118

61

Figure 4.8: Herd structure Alale division of West Pokot County

4.2.6.2 Prevalence of subclinical mastitis at animal (camel) level Alale division

At the animal (camel) level the prevalence of subclinical mastitis was 55.6% (5/9). In

terms of quarters affected, 11.1% (1/9) had only one quarter affected, 22.2% (2/9) had two

quarters affected, none of the camels had three quarters affected and another 22.2% (2/9) had all

the four quarters affected. From the results majority 44.4% (4/9) had both two and four quarters

infected.

4.2.6.3 Prevalence of subclinical mastitis at quarter level in Alale division

Of the 36 quarter samples cultured from the division 36.1% (13/36) quarter milk samples were

positive for subclinical mastitis while 63.9% (23/36) were negative. The right hind quarter was

Lactating camels16%

Dry camels38%

Breeding bulls3%

Immature males11%

Immature females

15%

Female calves11%

Male calves6%

Herd Structure Alale division

62

the most affected 44.4% (4/9) and the other quarters were equally 33.3% (3/9) infected. The

overall quarter-level prevalence of subclinical mastitis in this division was 36.1% (13/36).

Table 4. 25: Prevalence of subclinical mastitis at quarter level in Alale division

Quarter Positive Negative Total Prevalence (%) RFQ 3 6 9 33.3 RHQ 4 5 9 44.4 LFQ 3 6 9 33.3 LHQ 3 6 9 33.3 Total 13 23 36

63

CHAPTER FIVE

5.1 DISCUSSION

The observed milking frequencies practiced by the Pokot herders in this study were in

agreement with earlier studies in Kenya by Schwartz and Dioli (1992) who reported that milking

frequencies depended on season; yield and stage of lactation; availability of alternative food and

sex and health of the calf. McGovern (1995) reported that during wet seasons the Pokot people

milked their camels up to 4 times a day (morning, noon, evening and night), but in the dry

seasons the frequency was reduced to a single milking at night only.

The high overall prevalence (44.5%) of bacterial isolates from apparently normal camel

milk samples indicated a high percentage of subclinical mastitis in camels in West Pokot County.

However, the findings from this study was consistent with the findings of Woubit et al. (2001)

who reported high prevalence (51%) of mastitis in dromedary camels in Borena areas of South-

Western Ethiopia.

The most predominant bacterium isolated from this study was Staphylococcus aureus

with prevalence of 36.0% followed by E. coli with prevalence of 27.2% and Streptococcus

agalactiae & Staphylococcus epidermidis at 9.6% prevalence each. This finding is in agreement

with other findings from Eastern Sudan (Obied et al., 1996), Ethiopia (Workneh et al., 2002;

Kerro and Tareke, 2003; Biffa et al,. 2005 and Almaw et al., 2008) and from Kenya (Younan et

al., 2001) who reported that Staphylococcus aureus and Streptococcus agalactiae were the most

common causes of camel mastitis. It has also been reported in Kenya (Maina, 1984, Omore,

1997) that Staphylococcus aureus was the major cause of subclinical mastitis in bovine (63%).

As was also described by Younan et al. (2001), the prevalence of Staphylococci varies according

64

to different studies, but there is nearly no publication on bacteriological hygiene of milk where

Staphylococci are not mentioned (Eberlein, 2007).

The prevalence of E. coli has been reported by other authors at between 1.0 and 17.3 % in

samples taken from healthy camels (El-Jakee, 1998; Abdel Gadir et al., 2005). Therefore the

prevalence of E. coli from this study was higher than what has been reported earlier in other

studies.

Barbour et al. (1985) and Younan, (2004) stressed that the mastitis in milking dromedary

camels caused by Staphylococcus aureus (Coagulase Positive) is not only of veterinary interest

but represents a direct threat to human health considering that S. aureus can produce heat stable

enterotoxins that are not inactivated during pasteurization of milk or production of milk products

and can provoke food intoxication (vomiting and diarrhoea). The Coagulase negative

Staphylococcus (CNS) most often isolated from camel milk is Staphylococcus epidermidis

(Tuteja et al., 2003; Abdel Gadir et al., 2005).

Escherichia coli is also of public health importance since it is a pathogenic bacterium that

can cause severe intestinal and extra-intestinal diseases in man (Kaper et al., 2004) as well as

mastitis in cows (Bradley & Green, 2001). Abdel Gadir et al. (2005) isolated E. coli mainly (99.0

% of the isolates) from camel quarters that showed signs of subclinical mastitis. They also

reported one case of clinical mastitis caused by E. coli. This pathogen is also a marker for fecal

contamination due to the fact that it is a commensal of the intestinal tract (Schmidt-Lorenz &

Spillmann, 1988). However, this holds true more for water than for food (Busse, 1985).

Almaw & Molla, (2000) and Younan, (2004) reported Streptococcus agalactiae as one of the

main causes of clinical mastitis in camels and a potential human pathogen, causing intestinal

infections mainly in newborns.

65

The results further showed that there was cross infection from different quarters of

individual camels and from camels within the same herd because identical pathogens were

isolated from them. The above suggested that there was strong probability of direct transmission

by the milkers from one camel to another through poor milking procedures. Since most of the

micro-organisms isolated (especially Staphylococcus spp.) are associated with clinical mastitis,

particular attention should be given to the management of lactating camels to avoid development

of clinical mastitis. Given that consumption of raw camel milk is a common practice among

camel keepers in Kenya, the results from this study indicated the importance of pasteurization (or

boiling) of camel milk before consumption since potentially infective bacterial agents were

isolated. The high prevalence of subclinical mastitis could be attributed to unhygienic milking

procedures in poor hygienic conditions of the milking areas and generally poor traditional

management practices, where housing and sanitation was not a major priority in the study area.

Earlier works (Birru, 1989; Girma, 2001 and Mungube et al., 2004) showed that animals were

much more infected by mastitis, mainly in areas where hygienic conditions were poor and

treatment of mastitis cases was not well pursued.

Most of the bacteria isolated in this study were gram-positive cocci. This finding is in

agreement with what was reported previously by Obied et al. (1996) and Woubit et al. (2001).

The results also indicated that the most affected quarters were the right quarters with high

prevalence of 48.4% in the right hind quarter (RHQ) and 45.3% in the right fore quarter (RFQ).

This suggested the likelihood that in most cases during milking, the calf was left to suckle the

left quarters and the right quarters were milked by the owners and because of poor hygienic

milking procedures the right quarters were at a higher risk of getting infected.

66

The results also showed that the Pokots commonly kept camels together with other

livestock such as cattle, sheep and goats (personal communication). During milking of the

livestock, the results showed that the camels were the last to be milked therefore there was a high

possibility of direct transmission of mastitis pathogens from other species of livestock to the

camel. It was also noticed during this study that camel herders (mostly young boys) kept on

milking camels throughout the day during grazing. This practice might have also contributed to

the higher transmission rates of camel mastitis among camels in the same herd. Frequent milking

and suckling by the calf keeps flushing the mastitis pathogens out but some studies have shown

that it is the major cause of mastitis in camels (Obeid et al., 1996; Abdurahman, 1996).

The results also showed that most Pokot pastoralists were using various traditional herbs to

manage camel mastitis. This is consistent with what has been observed elsewhere as reported by

Bornstein, (1993) and Hussein, (1993) who reported that pastoralists use various traditional

(ethno-veterinary) practices to treat sick camels in Ethiopia.

67

CHAPTER SIX

6.1 CONCLUSSIONS AND RECOMMENDATIONS

6.1.1 CONCLUSSIONS

• The fact that the pathogens isolated from camel milk samples in this study were bacteria

that cause both environmental and contagious mastitis indicated that proper management

of lactating camels and adequate hygienic conditions of the environment are required in

order to minimize occurrence of mastitis in the study areas.

• It can also be concluded from the results of this study that mastitis was prevalent in

camels in West Pokot and was a serious problem that affected camels which are essential

for livelihoods of many nomadic tribes that live in the ASALs.

• More efforts are required to improve the general udder health in order to prevent and

control subclinical mastitis in camels.

• Also camel producers need to be trained or capacity built on the importance of hygienic

milking practices in order to minimize the potential adverse effect of mastitis on the yield

and quality of camel milk.

• The most important way to continuously produce camel milk of good quality is to keep

the mastitis level in the herd under optimum control.

68

6.1.2 RECOMMENDATIONS

• Camel producers and any other camel milk consumers should avoid consuming raw

camel milk but instead boil the milk before consuming.

• Hygienic milking procedures should be followed when milking camels.

• Milking order where you milk non mastitic camels first and camels or quarters with

mastitis infections last should be adhered to.

• Treatment of camels with mastitis infections using the conventional drugs should be

promoted while avoiding non-conventional treatment.

• There is need to create awareness on camel mastitis among camel keepers. At the

moment there is low level of awareness among pastoralists.

• More veterinary extension staff should be trained on camel mastitis diagnosis and control

as it affects camel productivity in the ASALs.

• Several mastitis control strategies should be put in place such as milking procedures,

milking order, strict hygiene, post milking teat disinfection, use of antibiotic dry-off

therapy and the culling of persistently infected camels.

69

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8.0 APPENDICES

8.1 APPENDIX I: A CROSS SECTIONAL STUDY QUESTIONNAIRE

Part A: Household Information

1) Interviewer ……………………………… Date…………………………….

(one form to be completed for each household)

2) Survey (household)…………………….. Division………………………

3) Location……………………………….. Sub location………………………

4) Owner’s name………………….. …………………….. Sex….. Age……

5) Interviewee……………………………..

6) Camel Herd composition:

a) Number of lactating camels…………..

b) Number of dry camels……………….

c) Breeding bulls…………………..

d) Immature males…………………

e) Immature females……………………..

f) Male calves…………………………..

g) Female calves………………………….

7) Camel milk production (per day);

87

a) Total milk produced………………..

b) Total milk consumed (domestic)……………………….

c) Total milk sold……………………………….

d) Price per Litre (Ksh)……………………………

8) Other Livestock owned by the family;

a) Cattle………….

b) Goats …………

c) Sheep ………….

d) Donkeys ……..

e) Others ………….

9) For how long have you been keeping camels?

a) < 1 year

b) ≥1year ≤ 5years

c) > 5years

10) Have you heard of mastitis in camel? Yes…… No……

11) What is the local name (Pokot) of mastitis……………………..

12) Have you ever had any case of mastitis? Yes ……. No ……

13) If yes what signs did you see to know it is mastitis?

a) Swollen and painful udder/quarter

b) Bloody milk

88

c) Reduced milk

d) Other signs………………………………

14) Was any treatment given? Yes….. No ……

15) Who administered the treatment?

a) Veterinary officer

b) Animal Health Assistant

c) Community Animal Health Worker

d) Myself

e) Others (specify)……………………………

16) Did the camel recover? Yes ….. No……

17) Who milks the camels?

a) Owner

b) Wife

c) Children

d) Other (specify)……………………………..

18) If you have more than one milking camels, when a camel has mastitis, do you milk it

last? Yes….. No…….

19) Are the camels milked using proper technique? (Observe)…………………………….

20) Do you prepare (wash) the udder before milking? Yes …… No……..

89

21) How often do you remove the manure from the boma where the milking camels spent in

the night?

a) < 3 months

b) ≥3 months ≤ 6 months

c) > 6 months ≤ 1 year

d) > 1 year.

22) What are the common camel diseases in the Manyatta?

a)………………………………

b)………………………………..

c)………………………………….

d)…………………………………..

e)……………………………………

Part B: Individual Camel Information

1) Camel’s Name……………….

(one form to be completed for each individual camel)

2) Camel’s breed…………………..

3) Parity ………………………..

4) Last calving date…………………….

5) Current milk production per day……………………..

6) Current number of functional teats (observe)……………………

7) Has the camel ever had mastitis during the current lactation? Yes …….No…….

8) Any abnormalities on the teats? (observe) Yes …….No………..

9) If yes describe the lesions…………………………………………….

90

10) Any abnormalities on the udder? (observe) Yes ……No……..

11) If yes describe the abnormalities………………………………………………

12) Milk samples taken from the quarters:

a) Right forequarter sample No……………

b) Left forequarter sample No……………..

c) Right hindquarter sample No…………….

d) Left hindquarter sample No……………….

13) Carry out CMT and indicate the results:

a) Right forequarter………………………………..

b) Left forequarter…………………………………….

c) Right hindquarter……………………………………….

d) Left hindquarter………………………………………..

14) Any other additional

information………………………………………………………………….

CMT (California Mastitis Test)

CMT is the most indirect test used to detect Subclinical Mastitis as the degree of Gel formation

is related with the number of cells in the milk.

Procedure: An equal volume of CMT Reagent and milk sample is mixed. The reactions are then

scored and interpreted as follows;

a) Score 1….. No reaction

b) Score 2……Slight slime which tends to disappear with continued swirling.

91

c) Score 3…….Distinct slime but without Gel formation.

d) Score 4……..Immediate formation of Gel which moves as a mass during

swirling.

e) Score 5…….The formed Gel develops a convex surface and also adheres to the

bottom of the paddle.

92

8.2 APPENIX II: Laboratory Results on bacterial culture and identification

HH.

No

Camel

No.

Parit

y

Lactation

Stage

RF

Q

RH

Q

LF

Q

LH

Q

Isolated Organism

(s)

REMARK

S

1 01 6th 2 months +ve -ve +ve +ve S. aureus Positive

02 3rd 6 months +ve -ve -ve +ve S. aureus Positive

03 4th 7 months +ve -ve -ve -ve S. aureus Positive

04 1st 6 months -ve +ve +ve -ve S. aureus Positive

05 2nd 3 months +ve -ve -ve -ve E. coli Positive

06 1st 6 months -ve -ve -ve +ve S. aureus Positive

2 07 3rd 1 month +ve -ve +ve +ve S. aureus Positive

08 3rd 4 months +ve +ve +ve +ve S. aureus Positive

3 09 3rd 7 months -ve -ve -ve +ve S. aureus &E. coli Positive

10 1st 7 months -ve -ve -ve -ve nil Negative

4 11 3rd 6 months -ve +ve -ve +ve S. aureus Positive

5 12 6th 3 months -ve +ve -ve +ve S. aureus &E. coli Positive

6 13 4th 4 months -ve -ve -ve +ve S. aureus &E. coli Positive

7 14 2nd 3 months +ve +ve -ve -ve S. aureus &E. coli Positive

15 1st 1 month +ve -ve +ve +ve S. aureus &E. coli Positive

8 16 3rd 5 months -ve -ve +ve -ve S. aureus Positive

17 2nd 6 months -ve -ve -ve -ve nil Negative

93

9 18 2nd 1 month +ve +ve +ve -ve E. coli Positive

10 19 1st 1 month -ve -ve -ve -ve nil Negative

11 20 3rd 3 days +ve -ve +ve -ve S. aureus &E. coli Positive

21 2nd 4 months +ve -ve -ve -ve E. coli Positive

22 3rd 4 months +ve +ve +ve +ve S. aureus &E. coli Positive

12 23 2nd 1 months +ve +ve -ve -ve S. aureus Positive

24 1st 1 month -ve -ve -ve +ve S. aureus Positive

13 25 3rd 1 month +ve +ve +ve -ve S. aureus &E. coli Positive

14 26 3rd 5 months -ve -ve -ve +ve E. coli Positive

15 27 4th 9 months -ve -ve +ve -ve S. aureus Positive

16 28 2nd 3 days -ve +ve -ve -ve E. coli Positive

17 29 1st 6 months -ve -ve -ve -ve nil Negative

18 30 2nd 9 months -ve -ve +ve -ve Micro-cocci Positive

19 31 3rd 6 months -ve +ve -ve -ve S. aureus Positive

20 32 4th 2 weeks -ve -ve -ve -ve nil Negative

21 33 2nd 3 months -ve +ve +ve -ve S. aureus Positive

22 34 2nd 4 months +ve +ve +ve +ve S. aureus Positive

35 3rd 4 months +ve +ve +ve -ve S. aureus &E. coli Positive

23 36 7th 4 months +ve +ve +ve +ve E. coli Positive

37 4th 3 months +ve +ve +ve +ve S. aureus Positive

24 38 2nd 1 month -ve +ve -ve -ve S. aureus Positive

39 1st 1 month -ve +ve -ve -ve Pseudomonas Positive

94

25 40 3rd 7 months -ve +ve +ve -ve S. aureus &E. coli Positive

26 41 3rd 1 month +ve +ve -ve +ve S. aureus Positive

27 42 1st 1 month -ve -ve -ve -ve nil Negative

28 43 3rd 3 months -ve -ve +ve +ve S. aureus Positive

29 44 1st 2 months +ve -ve +ve +ve S. aureus &E. coli Positive

30 45 4th 3 months +ve +ve +ve +ve S. aureus &E. coli Positive

31 46 2nd 1 month -ve -ve -ve -ve nil Negative

32 47 3rd 10

months

+ve -ve +ve +ve S. epidermidis, S.

agalactiae & E. coli

Positive

48 4th 12

months

+ve +ve +ve +ve S. agalactiae & E.

coli

Positive

49 5th 11

months

+ve -ve -ve +ve S. epidermidis, & E.

coli

Positive

33 50 1st 16

months

+ve +ve +ve -ve S. epidermidis, S.

agalactiae & E. coli

Positive

51 1st 8 months +ve -ve -ve +ve S. epidermidis, S.

agalactiae & E. coli

Positive

34 52 5th 10

months

+ve +ve +ve +ve S. epidermidis, S.

agalactiae & E. coli

Positive

35 53 6th 1 week +ve +ve +ve -ve S. aureus & E. coli Positive

54 4th 1 month -ve -ve -ve -ve nil Negative

55 2nd 1 month -ve -ve -ve -ve nil Negative

95

36 56 4th 4 months -ve +ve +ve +ve S. aureus & S.

agalactiae

Positive

57 2nd 11

months

-ve +ve -ve -ve S. epidermidis, S.

agalactiae & E. coli

Positive

58 5th 4 months -ve +ve -ve -ve S. aureus Positive

37 59 3rd 5 months +ve +ve +ve -ve S. aureus & S.

agalactiae

Positive

60 5th 5 months -ve +ve -ve -ve S. agalactiae Positive

38 61 2nd 9 months -ve -ve -ve -ve nil Negative

39 62 5th 3 months +ve -ve +ve +ve S. aureus & S.

agalactiae

Positive

40 63 2nd 2 weeks -ve -ve -ve -ve nil Negative

64 4th 14

months

+ve +ve +ve +ve S. aureus &E. coli Positive

65 2nd 13

months

-ve -ve -ve -ve nil Negative

41 66 2nd 9 months -ve +ve +ve -ve S. aureus, S.

epidermidis & E.

coli

Positive

67 3rd 8 months +ve +ve +ve +ve S. aureus & S.

epidermidis

Positive

68 1st 2 months -ve +ve -ve -ve E. coli Positive

96

42 69 2nd 5 months -ve -ve -ve -ve nil Negative

43 70 1st 2 months -ve -ve -ve -ve nil Negative

71 3rd 3 months +ve -ve -ve +ve S. aureus & S.

epidermidis

Positive

44 72 6th 2 days -ve -ve -ve -ve nil Negative

73 2nd 9 months -ve -ve -ve -ve nil Negative

74 1st 5 months +ve +ve +ve +ve S. aureus & S.

agalactiae

Positive

75 2nd 8 months +ve +ve +ve +ve S. aureus &E. coli Positive

76 1st 4 months -ve +ve +ve +ve S. aureus Positive

77 2nd 12

months

+ve +ve -ve -ve S. aureus Positive

45 78 4th 9 months -ve -ve -ve +ve E. coli Positive

79 2nd 10

months

+ve +ve +ve +ve S. agalactiae & S.

aureus

Positive

80 2nd 8 months -ve +ve -ve +ve S. agalactiae & E.

coli

Positive

81 5th 8 months -ve -ve -ve -ve nil Negative

82 1st 6 months +ve +ve -ve -ve S. aureus &E. coli Positive

83 4th 8 months -ve -ve -ve -ve nil Negative

84 3rd 9 months - ve -ve +ve -ve S. epidermidis Positive

46 85 7th 8 months -ve -ve -ve -ve nil Negative

97

86 5th 5 month +ve -ve -ve +ve S. aureus Positive

87 2nd 4 months +ve +ve +ve +ve S. epidermidis Positive

88 4th 6 months -ve -ve -ve -ve nil Negative

89 1st 7 months -ve -ve -ve -ve nil Negative

47 90 5th 6 months +ve +ve -ve -ve S. aureus &E. coli Positive

48 91 3rd 15

months

+ve +ve +ve -ve S. aureus & S.

epidermis

Positive

49 92 1st 5 months -ve +ve -ve +ve E. coli Positive

50 93 3rd 2 months -ve +ve +ve +ve S. epidermidis & E.

coli

Positive

51 94 2nd 2 months +ve -ve -ve -ve E. coli Positive

52 95 1st 2 months -ve -ve -ve -ve nil Negative

98